Disalt inhibitors of il-12  production

ABSTRACT

This invention relates to disalt nitrogen-heteroaryl inhibitors of IL-12 production, and related methods and pharmaceutical compositions.

RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.60/562,150, filed on Apr. 13, 2004, the entire teachings of which areincorporated herein by reference.

TECHNICAL FIELD

This invention relates to disalt inhibitors of IL-12 production, andrelated methods and pharmaceutical compositions.

BACKGROUND

Interleukin-12 (IL-12) is a heterodimeric cytokine (p70) which plays keyroles in immune responses by bridging innate resistance andantigen-specific adaptive immunity. Trinchieri (1993) Immunol Today 14:335. For example, it promotes type 1 T helper cell (T_(H)1) responsesand, hence, cell-mediated immunity. Chan et al. (1991) J Exp Med 173:869; Seder et al. (1993) Proc Natl Acad Sci USA 90: 10188; Manetti etal. (1993) J Exp Med 177: 1199; and Hsieh et al. (1993) Science 260:547. Interleukin-12 (IL-12) is a di-sulfide linked heterodimericcytokine composed of two independently regulated subunits, p35 and p40.IL-12 is produced by phagocytic cells and antigen presenting cells, inparticular, macrophages and dendritic cells, upon stimulation withbacteria, bacterial products such as lipopolysaccharide (LPS), andintracellular parasites. The well-documented biological functions ofIL-12 are induction of interferon-γ expression from T and NK cells anddifferentiation toward the T_(H)1 T lymphocyte type. IFN-γ, expressionof which is induced by IL-12, is a strong and selective enhancer ofIL-12 production from monocytes and macrophages. The cytokine IL-23 is aheterodimer composed of a p19 subunit and the same p40 subunit of IL-12.IL-23, similarly to IL-12, is involved in type 1 immune defenses andinduces IFN-γ secretion from T cells. IL-27 is formed by the associationof EBI3, a polypeptide related to the p40 subunit of IL-12, and p28, aprotein related to the p35 subunit of IL-12. IL-27 promotes the growthof T cells and is thought to play a role in the differentiation ofT_(H)1 cells. Pflanz et al., Immunity (2002), 16:779-790.

It has been suggested that, particularly in chronic diseases in whichthere is ongoing production of IFN-γ, IL-12 production is augmented byIFN-γ. It is presumed that after an infective or inflammatory stimulusthat provokes IL-12 production, the powerful feedback loop promotesIL-12- and IL-23-induced IFN-γ to further augment IL-12 production,leading to consequent excessive production of pro-inflammatorycytokines. Furthermore, it has been suggested that IL-27 induces theexpression of T-bet, a major T_(H)1-specific transcription factor, andit's downstream target IL-12R β2, independently of IFN-γ. In addition,IL-27 suppresses the expression of GATA-3. GATA-3 inhibits T_(H)1development and causes loss of IL-12 signaling through suppression ofIL-12R β2 and Stat4 expression. Lucas et al., PNAS (2003),100:15047-15052.

IL-12 plays a critical role in multiple-T_(H)1 dominant autoimmunediseases including, but not limited to, multiple sclerosis, sepsis,myasthenia gravis, autoimmune neuropathies, Guillain-Barré syndrome,autoimmune uveitis, autoimmune hemolytic anemia, pernicious anemia,autoimmune thrombocytopenia, temporal arteritis, anti-phospholipidsyndrome, vasculitides, Wegener's granulomatosis, Behcet's disease,psoriasis, psoriatic arthritis, dermatitis herpetiformis, pemphigusvulgaris, vitiligo, Crohn's disease, ulcerative colitis, interstitialpulmonary fibrosis, myelofibrosis, hepatic fibrosis, myocarditis,thyroditis, primary biliary cirrhosis, autoimmune hepatitis, Type 1 orimmune-mediated diabetes mellitus, Grave's disease, Hashimoto'sthyroiditis, autoimmune oophoritis and orchitis, autoimmune disease ofthe adrenal gland; rheumatoid arthritis, juvenile rheumatoid arthritis,systemic lupus erythematosus, scleroderma, polymyositis,dermatomyositis, spondyloarthropathies, ankylosing spondylitis,Sjogren's syndrome and graft-versus-host disease. See, for example,Gately et al. (1998) Annu Rev Immunol. 16: 495; and Abbas et al. (1996)Nature 383: 787.

Inhibiting IL-12 overproduction, or inhibiting the production ofcytokines such as IL-23 and IL-27 which promote IL-12 production and/orT_(H)1 development is an approach to treating the just-mentioneddiseases. Trembleau et al. (1995) Immmunol. Today 16: 383; and Adoriniet al. (1997) Chem. Immunol. 68: 175. For example, overproduction ofIL-12 and the resultant excessive T_(H)1 type responses can besuppressed by modulating IL-12, IL-23 and/or IL-27 production.Therefore, compounds that down-regulate IL-12, IL-23 and/or IL-27production can be used for treating inflammatory diseases. Ma et al.(1998) Eur Cytokine Netw 9: 54.

IL-12 also plays a role in bone loss diseases, particularly thoseinvolving osteoclasts. Osteoclasts are unique multinucleated cellswithin bone that are responsible for bone degradation and resorption.These are the only cells in the body known to be capable of thisfunction. Osteoclasts have a high capacity for the synthesis and storageof enzymes, including acid hydrolases and carbonic anhydrase isoenzymeII. Osteoclasts share phenotypic characteristics with circulatingmonocytes and tissue macrophages (N. Kurihara et al., Endocrinology 126:2733-41 (1990); G. Hattersley et al, Endocrinology 128: 259-62 (1991)).These cells are derived from mononuclear precursors that are the progenyof stem-cell populations located in the bone marrow, spleen, and liver.Proliferation of these stem-cell populations produces osteoclasticprecursors, which migrate via vascular routes to skeletal sites. Thesecells then differentiate and fuse with each other to form osteoclasts,or alternatively, fuse with existing osteoclasts.

The regulation of osteoclastic formation and activity is only partlyunderstood but it is known that excessive bone resorption by osteoclastscontributes to the pathology of many human diseases associated withexcessive bone loss, including periodontal disease, non-malignant bonedisorders (such as osteoporosis, Paget's disease of bone, osteogenesisimperfecta, fibrous dysplasia, and primary hyperparathyroidism) estrogendeficiency, inflammatory bone loss, bone malignancy, arthritis,osteopetrosis, and certain cancer-related disorders (such ashypercalcemia of malignancy (HCM), osteolytic bone lesions of multiplemyeloma and osteolytic bone metastases of breast cancer and othermetastatic cancers).

SUMMARY

This invention relates to disalt nitrogen-heteroaryl inhibitors of IL-12production, and related methods of making and using such compounds, andpharmaceutical compositions thereof.

In one aspect, the invention relates to a disalt represented by formula(I):

or a pharmaceutically acceptable solvate, clathrate, or prodrug thereof,wherein:

R¹ is —X

[N

]_(t)B;

X is C(R^(c)), O, S, S(O), S(O₂), or NR^(c);

t is 0 or 1;

each of

and

is, independently a single or double bond;

B is substituted or unsubstituted aryl, substituted or unsubstitutedheteroaryl, C(R^(a))(R^(b)), or L-B′;

each of R₂ and R₄, independently, is R^(c), halogen, nitro, cyano,azide, isothionitro, SR^(c), or OR^(c); or R₂ and R₄, taken together, is═O;

R₃ is R^(c), halogen, CN, alkenyl, alkynyl, OR^(c), OC(O)R^(c),SO₂R^(c), S(O)R^(c), S(O₂)NR^(c)R^(d), SR^(c), NR^(c)R^(d),NR^(c)COR^(d), NR^(c)(O)OR^(d), NR^(c)(O)NR^(c)R^(d), NR^(c)SO₂R^(d),COR^(c), C(O)OR^(c), or C(O)NR^(c)R^(d);

R₆ is H, alkyl, R^(c), halogen, nitro, cyano, azide, isothionitro,SR^(c), or OR^(c);

n is 0, 1, 2, 3, 4, 5, 6, or 7;

Y is a covalent bond, CH₂, C(O), C═N—R^(c), C═N—OR^(c), C═N—SR^(c), O,S, S(O), S(O)₂, or NR^(c);

Z is N or CH;

each of Q, U and V is, independently, N or CR^(c), provided that atleast one of Q, U, and V is N;

W is O, S, S(O), S(O)₂, NR^(c), or NC(O)R^(c);

L is O, S, S(O), S(O)₂, C(CR^(c))₂, or NR^(c);

B′ is substituted or unsubstituted aryl or substituted or unsubstitutedheteroaryl;

each of R^(a) and R^(b) is, independently, hydrogen, an optionallysubstituted alkyl, an optionally substituted aryl, or an optionallysubstituted heteroaryl;

each of R^(c) and R^(d), independently, is H, an optionally substitutedalkyl, an optionally substituted aryl, an optionally substitutedaralkyl, an optionally substituted heteroaryl, an optionally substitutedheteroaralkyl, an optionally substituted cyclyl, an optionallysubstituted heterocyclyl, or an alkylcarbonyl; and

each M⁻ is a conjugate base of a Bronsted acid.

In one embodiment, the invention relates to a disalt represented byformula (I), wherein each of Q, U and V is, independently, N or CH,provided that at least one of Q, U, and V is N.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein Q, U, and V each are N.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein two of Q, U and V are N, and the other is CH.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein Q and U each are N and V is CH.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein U and V are N, and Q is CH.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein Q and V are N and U is CH.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein one of Q, U and V is N, and the other two are eachCH.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein U is N and Q and V each are CH.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein Q is N and U and V each are CH.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein V is N and Q and U each are CH.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein Y is a covalent bond, O, S, NH or CH₂, and n is 0,1, 2, 3, or 4.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein R₂ and R₄ each are hydrogen.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein R₃ is optionally substituted aryl or optionallysubstituted heteroaryl.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein R₃ is optionally substituted heteroaryl.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein R₃ is pyridinyl.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein R₃ is OR^(c), SR^(c), C(O)OR^(c), NR^(c)R^(d), orC(O)NR^(c)R^(d).

In another embodiment, the invention relates to a disalt represented byformula (I) wherein R₃ is optionally substituted heterocyclyl.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein R₃ is morpholino.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein R³ is heteroaryl, heterocyclyl, or NR^(c)R^(d),wherein each of R^(c) and R^(d) of NR^(c)R^(d) is, independently,hydrogen, alkyl, cyclyl, or heterocyclyl.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein R₃ is

wherein:

each of A and A′, independently, is O, S, S(O), S(O)₂, or NH;

each of R₁₃ and R₁₄, independently is H, optionally substituted alkyl,optionally substituted aryl, or optionally substituted heteroaryl; and

m is 1 or 2.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein Z is N and W is O.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein t is 0, and

is a single bond.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein X is —NH— or —N(CH₃)—.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein t is 0,

is a single bond, and X is —NH— or —N(CH₃)—.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein B is:

wherein:

D is O, S, S(O), S(O)₂, or NR^(m);

R^(j), for each occurrence, is independently, halogen, CN, hydroxyl,alkyl, optionally substituted aryl, optionally substituted heteroaryl,alkoxyl, optionally substituted aryloxyl, or optionally substitutedheteroaryloxyl; or two R^(j) attached to two consecutive carbonstogether form a fused benzene ring;

R^(m) is H, alkyl, or alkylcarbonyl; and

r is 0, 1, or 2.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein B is optionally substituted aryl.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein B is naphthyl or

wherein:

R^(g) is H, halogen, CN, alkyl, cyclyl, alkyloxy, alkylcarbonyl,alkyloxycarbonyl, aryloxycarbonyl, heteroaryloxycarbonyl, hydroxyalkyl,alkylamino, or alkylaminocarbonyl;

R^(h), for each occurrence, is independently, halogen, NO₂, CN, alkyl,aryl, heteroaryl, OR^(c), OC(O)R^(c), SO₂R^(c), S(O)R^(c),S(O)₂NR^(c)R^(d), SR^(c), NR^(c)R^(d), NR^(c)C(O)OR^(d),NR^(c)C(O)OR^(d), NR^(c)C(O)NR^(c)R^(d), NR^(c)C(NR)NR^(c)R^(d),NR^(c)SO₂R^(d), C(O)R^(c), OC(O)R^(c), C(O)OR^(c), or C(O)NR^(c)R^(d);

R is an alkyl, an aryl, an aralkyl, —C(O)R^(c), —OR^(c), —SR^(c),—NR^(c)R^(d), hydroxylalkyl, nitro, cyano, haloalkyl, aminoalkyl, or—S(O)₂R^(c);

q is 0, 1, 2, 3, or 4.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein R³ is heteroaryl, heterocyclyl, or NR^(c)R^(d),wherein each of R^(c) and R^(d) of NR^(c)R^(d) is, independently,hydrogen, alkyl, cyclyl, or heterocyclyl.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein t is 1.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein X is NR^(c),

is a single bond,

is a double bond, t is 1, and B is C(R^(a))(R^(b)).

In another embodiment, the invention relates to a disalt represented byformula (I) wherein X is —NH— or —N(CH₃)—.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein one of R^(a) and R^(b) is

wherein:

E is NR^(i), O, S, S(O), or S(O)₂;

E′ is N or CR^(i);

R^(i) is H, alkyl, or alkylcarbonyl;

R^(g), R^(h), p and q are defined as above.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein one of R^(a) and R^(b) is

and the other of R^(a) and R^(b) is H or alkyl, wherein:

R^(g), R^(h), R^(i), and q are as defined above. In another aspect ofthis embodiment, R^(g) is H, methyl, ethyl, propyl, cyclopropyl,methoxy, ethoxy, methoxycarbonyl, or halogen; R^(h) is F, Cl, CN,methyl, methoxy, ethoxy, OC(O)CH₃, OC(O)C₂H₅, C(O)OH, C(O)OC₂H₅,C(O)NH₂, NHC(O)CH₃, or S(O)₂NH₂; R^(i) is H, methyl, ethyl, or acetyl;and q is 0, 1, or 2.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein one of R^(a) and R^(b) is:

and the other of R^(a) and R^(b) is H or alkyl, wherein R^(g) is definedas above. In another aspect of this embodiment, R^(g) is halogen, analkyl or an alkyloxycarbonyl.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein X is C(R^(c)),

is a double bond,

is a single bond, and B is L-B′.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein L is —NH— or —N(CH₃)—.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein X is CH;

In another embodiment, the invention relates to a disalt represented byformula (I) wherein B′ is:

wherein E, E′, R^(g), R^(h), p, and q are defined as above.

In another embodiment, the invention relates to a disalt represented byformula (I) wherein B′ is

wherein R^(g), R^(h), and q are defined as above.

One or more embodiments of for the compounds represented by formulas (I)may be combined to form additional compounds of the invention. All suchcombinations are expressly encompassed in this invention.

In another aspect, the invention relates to a disalt represented byformula (II):

or a pharmaceutically acceptable solvate, clathrate, or prodrug thereof,wherein B′, M⁻, Q, U, W, Y, Z, R₂, R₃, R₄, R₆, and n are defined asabove; and wherein:

G is O, S, S(O), S(O)₂, or NR^(e);

L₁ is O, S, S(O), S(O)₂, NR^(e), or C(O);

J is N or CR^(f);

R^(e), for each occurrence, is independently, H, alkyl, aryl, acyl,arylsulfonyl, or alkylsulfonyl; and

R^(f) is H, alkyl, aryl, acyl, arylsulfonyl, alkylsulfonyl, alkoxyl,amino, ester, amide, CN, or halogen.

In one embodiment, the invention relates to a disalt represented byformula (II) wherein G is NR^(e), and J is N.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein Z is N.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein W is O.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein L₁ is NR^(e).

In another embodiment, the invention relates to a disalt represented byformula (II) wherein each of Q and U is N.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein R₃ is a halogen, CN, an alkyl, an aryl, aheteroaryl, OR^(c), OC(O)R^(c), NR^(c)R^(d), NR^(c)C(O)R^(d),C(O)OR^(c), or C(O)NR^(d)R^(d).

In another embodiment, the invention relates to a disalt represented byformula (II) wherein R₃ is optionally substituted aryl, optionallysubstituted heteroaryl, optionally substituted aryloxyl, or optionallysubstituted heteroaryloxyl.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein R₃ is optionally substituted heteroaryl.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein R₃ is pyridinyl, triazolyl, tetrazolyl,pyrimidinyl, thiazolyl, indolyl, or indolizinyl.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein the compound is an N-oxide.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein B′ is an optionally substituted aryl.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein Y is NR^(c).

In another embodiment, the invention relates to a disalt represented byformula (II) wherein Y is O.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein B′ is

wherein R^(g), R^(h), and q are defined as above.

In another embodiment, the invention relates to a disalt represented byformula (II) wherein B′ is

wherein R^(g), R^(h), and q are defined as above. In another aspect ofthis embodiment, R^(g) is H, F, Cl, Br, I, CN, Me, Et, Pr, i-Pr, OMe, orOEt.

One or more embodiments of for the compounds represented by formulas(II) may be combined to form additional compounds of the invention. Allsuch combinations are expressly encompassed in this invention.

In another aspect, the invention relates to a disalt represented byformula (III):

or a pharmaceutically acceptable solvate, clathrate, or prodrug thereof,wherein:

each of R₇, R₈, R₉, and R₁₀, independently, is R^(c), halogen, CN,alkenyl, alkynyl, OR^(c), OC(O)R^(c), SO₂R^(c), S(O)R^(c),S(O₂)NR^(c)R^(d), SR^(c), NR^(c)R^(d), NR^(c)COR^(d), NR^(c)C(O)OR^(d),NR^(c)C(O)NR^(c)R^(d), NR^(c)SO₂R^(d), COR^(c), C(O)OR^(c), orC(O)NR^(c)R^(d);

one of R₁₁ and R₁₂ is -L₂-R₅ and the other is a group represented by thefollowing structural formula:

R₅ is an optionally substituted aryl, an optionally substitutedheteroaryl or a group represented by the following formula:

L₂ is O, S, S(O), S(O)₂, or NR^(c); and

M⁻, W, Z, R^(a), R^(b), R^(c), R^(d), and R₆ are defined as above.

In one embodiment, the invention relates to a disalt represented byformula (III) wherein R₅ is represented by the following formula:

In another embodiment, the invention relates to a disalt represented byformula (III) wherein L₂ is —NH—, —N(CH₃)—, —N(CH₂CH₃)—, or—N(C(O)CH₃)—.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein Z is N and W is O.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein one of R^(a) and R^(b) is an optionallysubstituted aryl or an optionally substituted heteroaryl, and the otherof R^(a) and R^(b) is H or alkyl.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein each of R₇, R₈, R₉, and R₁₀, independently, is H,—OH, an alkoxy, or an alkylcarbonyl.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein one of R^(a) and R^(b) is

wherein E, E′, R^(g), R^(h), p and q are defined as above.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein one of R^(a) and R^(b) is a group represented bythe following structural formula:

wherein R^(g), R^(h) and q are defined as above; and the other of R^(a)and R^(b) is H or alkyl. In another aspect of this embodiment, R^(g) isH, methyl, ethyl, propyl, cyclopropyl, methoxy, or ethoxy; and R^(h),for each occurrence, is independently, F, Cl, CN, methyl, methoxy,ethoxy, OC(O)CH₃, OC(O)C₂H₅, C(O)OH, C(O)OC₂H₅, C(O)NH₂, NHC(O)CH₃, orS(O₂)NH₂. In another aspect of this embodiment, R^(g) is methyl ormethoxy.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein Z is N and W is O.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein L₂ is —NH—, —N(CH₃)—, —N(CH₂CH₃)—, or —N(C(O)CH₃).

In another embodiment, the invention relates to a disalt represented byformula (III) wherein each of R₇, R₈, R₉, and R₁₀, independently, is H,—OH, an alkoxy, or an alkylcarbonyl.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein each of R₇ and R₁₀ is H and each of R₈ and R₉ isOCH₃.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein R₆ is H.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein R₁₂ is a group represented by the followingstructural formula:

and

R₁₁ is a group represented by the following formula:

In another embodiment, the invention relates to a disalt represented byformula (III) wherein R₁₂ is a group represented by the followingstructural formula:

and

R₁₁ is a group represented by the following structural formula:

In another embodiment, the invention relates to a disalt represented byformula (III) wherein R₅ is an optionally substituted aryl or anoptionally substituted heteroaryl.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein R₅ is:

wherein D, R^(j), and r are defined as above. In one aspect of thisembodiment, r is 1 or 2; and each R^(j) is, independently, methyl,ethyl, or propyl; or two R^(j) attached to two consecutive carbonstogether form a fused benzene ring.

In another embodiment, the invention relates to a disalt represented byformula (III) wherein R₅ is a group represented by the followingstructural formula:

In another embodiment, the invention relates to a disalt represented byformula (III) wherein each of R₇, R₈, R₉, and R₁₀, independent, is H orOR^(c).

In another embodiment, the invention relates to a disalt represented byformula (III) wherein R₁₂ is a group represented by the followingstructural formula:

and

R₁₁ is a group represented by the following structural formula:

In another embodiment, the invention relates to a disalt represented byformula (III) wherein R₁₂ is a group represented by the followingstructural formula:

and

R₁₁ is a group represented by the following structural formula:

One or more embodiments of for the compounds represented by formulas(III) may be combined to form additional compounds of the invention. Allsuch combinations are expressly encompassed in this invention.

In another embodiment, the invention relates to a disalt represented byformula (I), (II), or (III) wherein M⁻ is the conjugate base of aBronsted acid having a pKa in the range of between about −15 and about5.

In another embodiment, the invention relates to a disalt represented byformula (I), (II), or (III) wherein M⁻ is the conjugate base of aBronsted acid having a pKa of at most about −6.

In another embodiment, the invention relates to a disalt represented byformula (I), (II), or (III) wherein M⁻ is the conjugate base of aBronsted acid having a pKa of at most about −1.

In another embodiment, the invention relates to a disalt represented byformula (I), (II), or (III) wherein M⁻ is the conjugate base of aBronsted acid having a pKa of at most about 1.

In another embodiment, the invention relates to a disalt represented byformula (I), (II), or (III) wherein M⁻ is methanesulfonate.

In another embodiment, the invention relates to a disalt represented byformula (I), (II), or (III) wherein M⁻ is bromide.

In another embodiment, the invention relates to a disalt represented byformula (I), (II), or (III) wherein M⁻ is chloride.

In another embodiment, the invention relates to a disalt represented byformula (I), (II), or (III) wherein M⁻ is any combination of theaforementioned conjugate bases of a Bronsted acid.

Another aspect of the invention is a product (e.g., compound orcomposition) made by a process as delineated herein. The process caninclude one or more chemical transformations, salt formation, or otherchemical process as delineated herein; and can include one or morereagents, intermediates, solvents, or conditions, as delineated herein.

In another aspect, the present invention features a method for treatingor preventing an IL-12 production-related disorder such as multiplesclerosis, sepsis, myasthenia gravis, autoimmune neuropathies,Guillain-Barrë syndrome, autoimmune uveitis, autoimmune hemolyticanemia, pernicious anemia, autoimmune thrombocytopenia, temporalarteritis, anti-phospholipid syndrome, vasculitides, Wegener'sgranulomatosis, Behcet's disease, psoriasis, psoriatic arthritis,dermatitis herpetiformis, pemphigus vulgaris, vitiligo, Crohn's disease,ulcerative colitis, interstitial pulmonary fibrosis, myelofibrosis,hepatic fibrosis, myocarditis, thyroditis, primary biliary cirrhosis,autoimmune hepatitis, immune-mediated diabetes mellitus, Grave'sdisease, Hashimoto's thyroiditis, autoimmune oophoritis and orchitis,autoimmune disease of the adrenal gland; rheumatoid arthritis, juvenilerheumatoid arthritis, systemic lupus erythematosus, scleroderma,polymyositis, dermatomyositis, spondyloarthropathies, ankylosingspondylitis, Sjogren's syndrome and graft-versus-host disease. Themethod involves administering to a subject (e.g., a human or an animal)in need of treatment for an IL-12 production-related disorder aneffective amount of one or more disalts described herein, or apharmaceutically acceptable solvate, clathrate, or prodrug thereof, or apharmaceutical composition comprising an effective amount of one or moredisalts described herein, or a pharmaceutically acceptable solvate,clathrate, or prodrug thereof. In one embodiment, the method involvestreating or preventing an IL-12 production-related disorder selectedfrom the group consisting of rheumatoid arthritis, sepsis, Crohn'sdisease, multiple sclerosis, psoriasis, or immune-mediated diabetesmellitus. The identification of a subject in need of treatment for anIL-12 production-related disorder can be in the judgment of the subjector a health professional and can be subjective (e.g., opinion) orobjective (e.g., measurable by a test or a diagnostic method).

In one aspect, this invention features a method for treating orpreventing disorders associated with excessive bone loss, e.g.,periodontal disease, non-malignant bone disorders (e.g., osteoporosis,Paget's disease of bone, osteogenesis imperfecta, fibrous dysplasia, andprimary hyperparathyroidism), estrogen deficiency, inflammatory boneloss, bone malignancy, arthritis, osteopetrosis, and certaincancer-related disorders (e.g., hypercalcemia of malignancy (HCM),osteolytic bone lesions of multiple myeloma and osteolytic bonemetastases of breast cancer and other metastatic cancers). The methodincludes administering to a subject (e.g., a human or an animal) in needthereof an effective amount of one or more disalts described herein or apharmaceutically acceptable solvate, clathrate, or prodrug thereof, or apharmaceutical composition comprising an effective amount of one or moredisalts described herein or a pharmaceutically acceptable solvate,clathrate, or prodrug thereof. The method can also include the step ofidentifying that the subject is in need of treatment of diseases ordisorders described above. The identification can be in the judgment ofa subject or a health professional and can be subjective (e.g., opinion)or objective (e.g., measurable by a test or a diagnostic method).

In another aspect, this invention features methods for inhibitingosteoclast formation in vitro or in vivo. The method includes contactinga pre-osteoclast cell (e.g., a cell capable of forming an osteoclastcell upon differentiation and/or fusion) with an effective amount of adisalt described herein or a pharmaceutically acceptable solvate,clathrate, or prodrug thereof or a pharmaceutical composition comprisingan effective amount of a disalt described herein or a pharmaceuticallyacceptable solvate, clathrate, or prodrug thereof.

In a further aspect, this invention features methods of treating orpreventing a disorder associated with excessive bone resorption byosteoclasts in a subject in need thereof. The method includesadministering to a subject (e.g., a human or an animal) in need thereofan effective amount of one or more disalts described herein or apharmaceutically acceptable solvate, clathrate, or prodrug thereof, or apharmaceutical composition comprising an effective amount of one or moredisalts described herein or a pharmaceutically acceptable solvate,clathrate, or prodrug thereof. The method can also include the step ofidentifying that the subject is in need of treatment of diseases ordisorders described above. The identification can be in the judgment ofa subject or a health professional and can be subjective (e.g., opinion)or objective (e.g., measurable by a test or a diagnostic method).

The disalts of this invention can include the diprotonated IL-12production inhibitor compounds themselves, as well as their prodrugs, ifapplicable. As used herein and unless otherwise indicated, the term“prodrug” means a derivative of a compound that can hydrolyze, oxidize,or otherwise react under biological conditions (in vitro or in vivo) toprovide a compound of this invention. Prodrugs may only become activeupon such reaction under biological conditions, or they may haveactivity in their unreacted forms. Examples of prodrugs contemplated inthis invention include, but are not limited to, analogs or derivativesof compounds of any one of the formulae disclosed herein that comprisebiohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzableesters, biohydrolyzable carbamates, biohydrolyzable carbonates,biohydrolyzable ureides, and biohydrolyzable phosphate analogues. Otherexamples of prodrugs include derivatives of compounds of any one of theformulae disclosed herein that comprise —NO, —NO₂, —ONO, or —ONO₂moieties. Prodrugs can typically be prepared using well-known methods,such as those described by 1 BURGER′S MEDICINAL CHEMISTRY AND DRUGDISCOVERY (1995) 172-178, 949-982 (Manfred E. Wolff ed., 5^(th) ed).

As used herein and unless otherwise indicated, the terms“biohydrolyzable amide”, “biohydrolyzable ester”, “biohydrolyzablecarbamate”, “biohydrolyzable carbonate”, “biohydrolyzable ureide” and“biohydrolyzable phosphate analogue” mean an amide, ester, carbamate,carbonate, ureide, or phosphate analogue, respectively, that either: 1)does not destroy the biological activity of the compound and confersupon that compound advantageous properties in vivo, such as uptake,duration of action, or onset of action; or 2) is itself biologicallyinactive but is converted in vivo to a biologically active compound.Examples of biohydrolyzable amides include, but are not limited to,lower alkyl amides, α-amino acid amides, alkoxyacyl amides, andalkylaminoalkylcarbonyl amides. Examples of biohydrolyzable estersinclude, but are not limited to, lower alkyl esters, alkoxyacyloxyesters, alkyl acylamino alkyl esters, and choline esters. Examples ofbiohydrolyzable carbamates include, but are not limited to, loweralkylamines, substituted ethylenediamines, aminoacids,hydroxyalkylamines, heterocyclic and heteroaromatic amines, andpolyether amines.

As used herein, the term “pharmaceutically acceptable solvate,” is asolvate formed from the association of one or more solvent molecules toone of the disalts of this invention. The term solvate includes hydrates(e.g., hemi-hydrate, mono-hydrate, dihydrate, trihydrate, tetrahydrate,and the like).

The disalts of this invention may contain one or more asymmetric centersand thus occur as racemates and racemic mixtures, single enantiomers,individual diastereomers and diastereomeric mixtures. All such isomericforms of these disalts are expressly included in the present invention.The disalts of this invention may also contain linkages (e.g.,carbon-carbon bonds) wherein bond rotation is restricted about thatparticular linkage, e.g. restriction resulting from the presence of aring or double bond. Accordingly, all cis/trans and E/Z isomers areexpressly included in the present invention. The disalts of thisinvention may also be represented in multiple tautomeric forms, in suchinstances, the invention expressly includes all tautomeric forms of thedisalts described herein, even though only a single tautomeric form maybe represented (e.g., alkylation of a ring system may result inalkylation at multiple sites, the invention expressly includes all suchreaction products). All such isomeric forms of such disalts areexpressly included in the present invention. All crystal forms andpolymorphs of the disalts described herein are expressly included in thepresent invention.

Further, the aforementioned aromatic ring nitrogen-containing compoundsalso include their N-oxides. The term “N-oxides” refers to one or morenitrogen atoms, when present in an aromatic ring nitrogen-containingcompound, are in N-oxide form, i.e., N→O.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable disalts. The term“stable”, as used herein, refers to disalts, which possess stabilitysufficient to allow manufacture and which maintains the integrity of thedisalt for a sufficient period of time to be useful for the purposesdetailed herein (e.g., treating IL-12 production-related disorders suchas rheumatoid arthritis, sepsis, Crohn's disease, multiple sclerosis,psoriasis, or insulin-dependent diabetes mellitus, treating orpreventing disorders associated with excessive bone loss, e.g.,periodontal disease, non-malignant bone disorders (e.g., osteoporosis,Paget's disease of bone, osteogenesis imperfecta, fibrous dysplasia, andprimary hyperparathyroidism) estrogen deficiency, inflammatory boneloss, bone malignancy, arthritis, osteopetrosis, and certaincancer-related disorders (e.g., hypercalcemia of malignancy (HCM),osteolytic bone lesions of multiple myeloma and osteolytic bonemetastases of breast cancer and other metastatic cancers), inhibitingosteoclast formation in vitro or in vivo, or treating or preventing adisorder associated with excessive bone resorption by osteoclasts.).

Also within the scope of this invention are a composition comprising apharmaceutically acceptable carrier and one or more of the disaltsdescribed above. The compositions may be used for treating an IL-12production-related disorder (e.g., rheumatoid arthritis, sepsis, Crohn'sdisease, multiple sclerosis, psoriasis, or insulin-dependent diabetesmellitus), treating or preventing disorders associated with excessivebone loss, e.g., periodontal disease, non-malignant bone disorders(e.g., osteoporosis, Paget's disease of bone, osteogenesis imperfecta,fibrous dysplasia, and primary hyperparathyroidism) estrogen deficiency,inflammatory bone loss, bone malignancy, arthritis, osteopetrosis, andcertain cancer-related disorders (e.g., hypercalcemia of malignancy(HCM), osteolytic bone lesions of multiple myeloma and osteolytic bonemetastases of breast cancer and other metastatic cancers), inhibitingosteoclast formation in vitro or in vivo, or treating or preventing adisorder associated with excessive bone resorption by osteoclasts).

In another aspect, this invention features a pharmaceutical compositionthat contains a pharmaceutically acceptable carrier and an effectiveamount of at least one of the disalts of this invention, or apharmaceutically acceptable solvate, clathrate, or prodrug thereof.These compositions may further include one or more additional activeagents. The compositions are useful for treating or preventing one ormore of the disorders delineated herein.

Also within the scope of the invention is the use of one or more disaltsdescribed herein, or a pharmaceutically acceptable solvate, clathrate,or prodrug thereof, or a pharmaceutical composition comprising aneffective amount of one or more disalts described herein, or apharmaceutically acceptable solvate, clathrate, or prodrug thereof, forthe manufacture of a medicament. The medicament may be used for treatingan IL-12 production-related disorder (e.g., rheumatoid arthritis,sepsis, Crohn's disease, multiple sclerosis, psoriasis, orinsulin-dependent diabetes mellitus), treating or preventing disordersassociated with excessive bone loss, e.g., periodontal disease,non-malignant bone disorders (e.g., osteoporosis, Paget's disease ofbone, osteogenesis imperfecta, fibrous dysplasia, and primaryhyperparathyroidism) estrogen deficiency, inflammatory bone loss, bonemalignancy, arthritis, osteopetrosis, and certain cancer-relateddisorders (e.g., hypercalcemia of malignancy (HCM), osteolytic bonelesions of multiple myeloma and osteolytic bone metastases of breastcancer and other metastatic cancers), inhibiting osteoclast formation invitro or in vivo, or treating or preventing a disorder associated withexcessive bone resorption by osteoclasts).

The invention also relates to a method of making a disalt describedherein. In one embodiment, the method comprises contacting with aBronsted acid having a pKa in the range of between about −15 and about5, a compound represented by formula (Ia), (IIa) or (IIIa):

wherein R₁, R₂, R₃, R₄, R₆, R₇, R₈, R₉, R₁₀, R₁₁, R₁₂, B′, G, J, L₁, Q,U, V, W, Y, and n are defined as above.

In one embodiment, the method includes taking any one of theintermediate compounds described herein and reacting it with one or morechemical reagents in one or more steps to produce a compound describedherein.

In some embodiments, the disalts have one or more properties that areenhanced relative to the corresponding monoprotonated compounds and/orthe unprotonated compounds, e.g., bioavailability, solubility, meltingpoint, and stability), and advantageously can have more optimalformulation properties relative to those of the monoprotonated compoundsand/or the unprotonated compounds.

Other features and advantages of the invention will be apparent from thedescription and from the claims.

DETAILED DESCRIPTION

As used herein, the term “alkyl” refers to a straight-chained orbranched hydrocarbon group containing 1 to 12 carbon atoms. Alkyl groupsmay be optionally substituted with one or more substituents. Examples ofalkyl groups include methyl, ethyl, n-propyl, isopropyl, tert-butyl, andn-pentyl.

As used herein, the term “alkoxy” or “alkyloxy” refers to an alkyl groupthat is linked to another moiety by an oxygen (i.e., —O-alkyl). Thealkyl portion of an alkoxy group may be optionally substituted.

The term “alkenyl” refers to an unsaturated hydrocarbon chain that maybe a straight chain or branched chain containing 2 to 12 carbon atomsand at least one carbon-carbon double bond. Alkenyl groups may beoptionally substituted with one or more substituents.

The term “alkynyl” refers to an unsaturated hydrocarbon chain that maybe a straight chain or branched chain, containing the 2 to 12 carbonatoms and at least one carbon-carbon triple bond. Alkynyl groups may beoptionally substituted with one or more substituents.

The sp² or sp carbons of an alkenyl group and an alkynyl group,respectively, may optionally be the point of attachment of the alkenylor alkynyl groups.

The term “aryl” refers to a hydrocarbon monocyclic, bicyclic ortricyclic ring system having at least one aromatic ring. Aryl groups maybe optionally substituted with one or more substituents. In oneembodiment, 0, 1, 2, 3, 4, 5 or 6 atoms of each ring of an aryl groupmay be substituted by a substituent. Examples of aryl groups includephenyl, naphthyl, anthracenyl, fluorenyl, indenyl, azulenyl, and thelike.

As used herein, the term “alkylene” refers to an alkyl group that hastwo points of attachment. The term “(C₁-C₆)alkylene” refers to analkylene group that has from one to six carbon atoms. Non-limitingexamples of alkylene groups include methylene (—CH₂—), ethylene(—CH₂CH₂—), n-propylene (—CH₂CH₂CH₂—), isopropylene (—CH₂CH(CH₃)—), andthe like.

As used herein, the term “aralkyl” means an aryl group that is attachedto another group by a (C₁-C₆)alkylene group. Aralkyl groups may beoptionally substituted, either on the aryl portion of the aralkyl groupor on the alkylene portion of the aralkyl group, with one or moresubstituent. Representative aralkyl groups include benzyl,2-phenyl-ethyl, naphth-3-yl-methyl and the like.

The term “heteroaryl” refers to a mono-, bi- or tri-cyclic ring systemhaving at least one aromatic ring, wherein a monocyclic heteroaryl has5-8 members, a bicyclic heteroaryl has 8-12 members, and a tricyclicheteroaryl has 11-14 member. A heteroaryl may have 1-4 ring heteroatomsif monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms iftricyclic, said heteroatoms selected from O, N, or S, and the remainderring atoms being carbon (with appropriate hydrogen atoms unlessotherwise indicated). Heteroaryl groups may be optionally substitutedwith one or more substituents. In one embodiment, 0, 1, 2, 3, or 4 atomsof each ring of a heteroaryl group may be substituted by a substituent.Examples of heteroaryl groups include pyridyl, 1-oxo-pyridyl, furanyl,benzo[1,3]dioxolyl, benzo[1,4]dioxinyl, thienyl, pyrrolyl, oxazolyl,oxadiazolyl, imidazolyl thiazolyl, isoxazolyl, quinolinyl, pyrazolyl,isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, triazolyl,thiadiazolyl, isoquinolinyl, indazolyl, benzoxazolyl, benzofuryl,indolizinyl, imidazopyridyl, tetrazolyl, benzimidazolyl, benzothiazolyl,benzothiadiazolyl, benzoxadiazolyl, indolyl, tetrahydroindolyl,azaindolyl, imidazopyridyl, quinazolinyl, purinyl,pyrrolo[2,3]pyrimidinyl, pyrazolo[3,4]pyrimidinyl, and benzo(b)thienyl,3H-thiazolo[2,3-c][1,2,4]thiadiazolyl,imidazo[1,2-d]-1,2,4-thiadiazolyl, imidazo[2,1-b]-1,3,4-thiadiazolyl,1H,2H-furo[3,4-d]-1,2,3-thiadiazolyl,1H-pyrazolo[5,1-c]-1,2,4-triazolyl, pyrrolo[3,4-d]-1,2,3-triazolyl,cyclopentatriazolyl, 3H-pyrrolo[3,4-c]isoxazolyl,1H,3H-pyrrolo[1,2-c]oxazolyl, pyrrolo[2,1b]oxazolyl, and the like.

As used herein, the term “heteroaralkyl” or “heteroarylalkyl” means aheteroaryl group that is attached to another group by a (C₁-C₆)alkylene.Heteroaralkyl groups may be optionally substituted, either on theheteroaryl portion of the heteroaralkyl group or on the alkylene portionof the heteroaralkyl group, with one or more substituent. Representativeheteroaralkyl groups include 2-(pyridin-4-yl)-propyl,2-(thien-3-yl)-ethyl, imidazol-4-yl-methyl and the like.

The term “cyclyl” refers to a partially or fully saturated non-aromaticmono-cyclic or bi-cyclic hydrocarbon ring system having from 3 to 14ring atoms. Exemplary cyclyl rings include cyclopropyl, cyclohexanyl,cyclopentanyl, cyclohexenyl, cyclohexadienyl, cyclopentenyl, and thelike.

The term “heterocyclyl” refers to a partially or fully saturatednon-aromatic mono-cyclic or bi-cyclic ring system having from 3 to 14ring atoms. A heterocyclyl ring contains one or more heteroatoms (e.g.,O, N, or S) as part of the ring system and the remainder being carbon.Exemplary heterocyclyl rings include piperidinyl, piperazinyl,morpholinyl, thiomorpholinyl, 1,4-oxazepanyl, pyranyl,2-oxo-1H-pyridinyl, and the like.

The term “mercapto” refers to —SH. The term “alkyl sulfanyl” refers to—S-alkyl. The term “aryl sulfanyl” refers to —S-aryl. The alkyl or arylportion of an alkyl sulfanyl or an aryl sulfanyl may be optionallysubstituted.

As used herein, the term “halogen” or “halo” means —F, —Cl, —Br or —I.

As used herein, the term “haloalkyl” means an alkyl group in which oneor more (including all) the hydrogen radicals are replaced by a halogroup, wherein each halo group is independently selected from —F, —Cl,—Br, and —I. The term “halomethyl” means a methyl in which one to threehydrogen radical(s) have been replaced by a halo group. Representativehaloalkyl groups include trifluoromethyl, bromomethyl,1,2-dichloroethyl, 4-iodobutyl, 2-fluoropentyl, and the like.

As used herein, the term “haloalkoxy” means an alkoxy group in which oneor more (including all) the hydrogen radicals are replaced by a halogroup, wherein each halo group is independently selected from —F, —Cl,—Br, and —I.

The term “amino” refers to —NH₂. The term “alkylamino” refers to anamino group in which one hydrogen radical has been replaced by an alkylgroup. The term “dialkylamino” refers to an amino group in which twohydrogen radicals have been replaced by two independently selected alkylgroups Likewise, the term “arylamino” refers to an amino group in whichone hydrogen radical has been replaced by an aryl group. The term“diarylamino” refers to an amino group in which two hydrogen radicalshave been replaced by two independently selected aryl groups.

The term “azide” refers to a group having the formula —N═N⁺═N⁻

The term “mercaptoalkyl” refers to an alkyl substituent which is furthersubstituted with one or more mercapto groups. The term “mercaptoalkoxy”refers to an alkoxy substituent which is further substituted with one ormore mercapto groups.

The term “hydroxyalkyl” or “hydroxylalkyl” refers to an alkylsubstituent which is further substituted with one or more hydroxylgroups.

The term “sulfonylalkyl” refers to an alkyl substituent which is furthersubstituted with one or more sulfonyl groups. The term “sulfonylaryl”refers to an aryl substituent which is further substituted with one ormore sulfonyl groups.

The term alkylcarbonyl refers to an —C(O)-alkyl. The term“alkylcarbonylalkyl” refers to an alkyl substituent which is furthersubstituted with —C(O)-alkyl. The alkyl or aryl portion of alkylamino,dialkylamino, aminoalkyl, mercaptoalkyl, mercaptoalkoxy, hydroxyalkyl,mercaptoalkoxy, sulfonylalkyl, sulfonylaryl, alkylcarbonyl, andalkylcarbonylalkyl may be optionally substituted with one or moresubstituents.

The term “ester” refers to a —C(O)O—R^(c); or, where a divalent group isindicated, an “ester” group is —C(O)O— or —OC(O)—. An “amido” is an—C(O)NH₂. A divalent “amide” group is indicated, the group is —C(O)N— or—NC(O)—.

An alkyl, alkoxy, alkyl sulfanyl, alkylamino, dialkylamino, alkylene,alkenyl, alkynyl, cyclyl, heterocyclyl, aryl, aralkyl, and heteroarylmentioned above can be substituted or unsubstituted moieties. The term“substituted” refers to one or more substituents (which may be the sameor different), each replacing a hydrogen atom. Suitable substituents foran alkyl, alkoxy, alkyl sulfanyl, alkylamino, dialkylamino, alkylene,alkenyl, alkynyl, cyclyl, heterocyclyl, aryl, aralkyl, and heteroarylgroups include any substituent which will form a stable compound of theinvention. Examples of substituents for an alkyl, alkoxy, alkylsulfanyl,alkylamino, dialkylamino, alkylene, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, heterocyclyl, aryl, aralkyl, heteroaryl, andheteroarylalkyl include an alkyl, alkoxy, alkyl sulfanyl, alkylamino,dialkylamino, an alkenyl, an alkynyl, an cycloalkyl, an cycloalkenyl, anheterocyclyl, an aryl, an heteroaryl, an aralkyl, an heteraralkyl, ahaloalkyl, halo, cyano, nitro, haloalkoxy, —OR₁₅, —SR₁₅, —NR₁₇R₁₈,—C(O)NR₁₇R₁₈, —NR₁₅C(O)R₁₆, —C(S)NR₁₇R₁₈, —NR₁₅C(S)R₁₆, —C(NR₁₉)NR₁₇R₁₈,—NR₁₅C(NR₁₉)R₁₆, —C(O)R₁₅, —C(S)R₁₅, —C(NR₁₉)R₁₅, —C(O)OR₁₅, —C(O)SR₁₅,—OC(O)R₁₅, —SC(O)R₁₅, —C(S)OR₁₅, —C(S)SR₁₅, —OC(S)R₁₅, —SC(S)R₁₅,—C(NR₁₉)OR₁₅, —C(NR₁₉)SR₁₅, —OC(NR₁₉)R₁₅, —SC(NR₁₉)R₁₅,—NR₁₅C(O)NR₁₇R₁₈, —NR₁₅C(S)NR₁₇R₁₈, —NR₁₅C(NR₁₉)NR₁₇R₁₈, —OC(O)NR₁₇R₁₈,—OC(S)NR₁₇R₁₈, —OC(NR₁₉)NR₁₇R₁₈, —SC(O)NR₁₇R₁₈, —SC(S)NR₁₇R₁₈,—SC(NR₁₉)NR₁₇R₁₈, —SC(NR₁₉)NR₁₇R₁₈, —NR₁₅C(O)OR₁₆, —NR₁₅C(S)OR₁₆,—NR₁₅C(NR₁₉)OR₁₆, —NR₁₅C(O)SR₁₆, —NR₁₅C(S)SR₁₆, —NR₁₅C(NR₁₉)SR₁₆,—S(O)_(p)OR₁₅, —OS(O)_(p)OR₁₅, —OS(O)_(p)R₁₅, —S(O)_(p)R₁₅,—S(O)_(p)NR₁₇R₁₈, —NR₁₅S(O)_(p)R₁₆, —P(O)(OR₁₅)₂, —OP(O)(OR₁₅)₂,—P(S)(OR₁₅)₂, —OP(S)(OR₁₅)₂, —SP(O)(OR₁₅)₂, —OP(O)(OR₁₅)(SR₁₆), or—P(O)(OR₁₅)(SR₁₆), wherein R₁₇ and R₁₈, for each occurrence are,independently, H, an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, an optionallysubstituted cyclyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl; or R₁₇ and R₁₈ taken together with the nitrogen to whichthey are attached is optionally substituted heterocyclyl or optionallysubstituted heteroaryl; and R₁₅ and R₁₆ for each occurrence are,independently, H, an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, an optionallysubstituted cyclyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl. R₁₉, for each occurrence, is independently H, anoptionally substituted alkyl, an optionally substituted cyclyl, anoptionally substituted heterocyclyl, an optionally substituted aryl, anoptionally substituted heteroaryl, an optionally substituted aralkyl, anoptionally substituted heteroaralkyl, —C(O)R^(c), —OR₁₅, —SR₁₅,—NR₁₇R₁₈, hydroxylalkyl, nitro, cyano, haloalkyl, aminoalkyl, or—S(O)₂R^(c).

In addition, an alkyl, alkylene and any saturated portion of a alkenyl,cyclyl, alkynyl, aralkyl, heterocyclyl and heteroaralkyl groups, mayalso be substituted with ═O, ═S, ═N—R₁₉.

When a heterocyclyl, heteroaryl, or heteroaralkyl group contains anitrogen atom, it may be substituted or unsubstituted. When a nitrogenatom in the aromatic ring of a heteroaryl group has a substituent thenitrogen may be a quaternary nitrogen.

In one embodiment, suitable substituents for an alkyl, alkoxy, alkylsulfanyl, alkylamino, dialkylamino, alkylene, alkenyl, alkynyl, cyclyl,heterocyclyl, aryl, aralkyl, and heteroaryl include, but are not limitedto, halogen (F, Cl, Br, or I), hydroxyl, amino, alkylamino, arylamino,dialkylamino, diarylamino, cyano, nitro, mercapto, carbonyl, carbamido,carbamyl, carboxyl, thioureido, thiocyanato, sulfoamido, alkyl, alkenyl,alkyloxy, aryl, heteroaryl, cyclyl, heterocyclyl, wherein alkyl,alkenyl, alkyloxy, aryl, heteroaryl cyclyl, and heterocyclyl areoptionally substituted with alkyl, aryl, heteroaryl, halogen, hydroxyl,mercapto, cyano, or nitro.

Choices and combinations of substituents and variables envisioned bythis invention are only those that result in the formation of stablecompounds. The term “stable”, as used herein, refers to compounds whichpossess stability sufficient to allow manufacture and which maintainsthe integrity of the compound for a sufficient period of time to beuseful for the purposes detailed herein (e.g., therapeutic orprophylactic administration to a subject). Typically, such compounds arestable at a temperature of 40° C. or less, in the absence of excessivemoisture, for at least one week. Such choices and combinations will beapparent to those of ordinary skill in the art and may be determinedwithout undue experimentation.

As used herein, the terms “animal”, “subject” and “patient”, include,but are not limited to, a cow, monkey, horse, sheep, pig, chicken,turkey, quail, cat, dog, mouse, rat, rabbit, guinea pig and human(preferably, a human).

As used herein, the term “lower” refers to a group having up to fouratoms. For example, a “lower alkyl” refers to an alkyl radical havingfrom 1 to 4 carbon atoms; a “lower alkenyl” or “lower alkynyl” refers toan alkenyl or alkynyl radical having from 2 to 4 carbon atoms,respectively; and a lower alkoxy refers to an alkoxy having 1 to 4carbon atoms.

As used herein, the term “pharmaceutically acceptable solvate,” is asolvate formed from the association of one or more solvent molecules toone of the compounds of formula (I), (II) or (III). The term solvateincludes hydrates (e.g., mono-hydrate, dihydrate, trihydrate,tetrahydrate, and the like).

As used herein, the term “clathrate” means a compound of the presentinvention in the form of a crystal lattice that contains spaces (e.g.,channels) that have a guest molecule (e.g., a solvent or water) trappedwithin.

As used herein, the term “pre-osteoclast cell” is a cell capable offorming an osteoclast cell upon differentiation and/or fusion andincludes without limitation, circulating monocytes and tissuemacrophages (N. Kurihara et al., Endocrinology 126: 2733-41 (1990)).Without wishing to be bound by theory, pre-osteoclasts are converted toactivated osteoclasts in a process thought to involve two factorsproduced by pre-osteoblasts, M-CSF and ODF. These factors activatecertain genes that are needed for the conversion of a pre-osteoclastinto an osteoclast.

The disalts described herein are useful to treat and prevent any IL-12production-related disorders, e.g., inflammatory disorders, immunediseases, neurological disorders and bone loss diseases.

The term “inflammatory disorders” includes any inflammatory disease,disorder or condition caused, exasperated or mediated by IL-12production. Such inflammatory disorders may include, without limitation,asthma, adult respiratory distress syndrome, systemic lupuserythematosus, inflammatory bowel disease (including Crohn's disease andulcerative colitis), multiple sclerosis, insulin-dependent diabetesmellitus, autoimmune arthritis (including rheumatoid arthritis, juvenilerheumatoid arthritis, psoriatic arthritis), inflammatory pulmonarysyndrome, pemphigus vulgaris, idiopathic thrombocytopenic purpura,autoimmune meningitis, myasthenia gravis, autoimmune thyroiditis,dermatitis (including atopic dermatitis and eczematous dermatitis),psoriasis, Sjogren's Syndrome (including keratoconjunctivitis siccasecondary to Sjogren's Syndrome), alopecia areata, allergic responsesdue to arthropod bite reactions, aphthous ulcer, iritis, conjunctivitis,keratoconjunctivitis, cutaneous lupus erythematosus, scleroderma,vaginitis, proctitis, drug eruptions (such as Stevens-Johnson syndrome),leprosy reversal reactions, erythema nodosum leprosum, autoimmuneuveitis, allergic encephalomyelitis, aplastic anemia, pure red cellanemia, idiopathic thrombocytopenia, polychondritis, Wegener'sgranulomatosis, chronic active hepatitis, Graves ophthalmopathy, primarybiliary cirrhosis, uveitis posterior and interstitial lung fibrosis.

“Inflammatory disorders” expressly include acute inflammatory disorders.Examples of acute inflammatory disorders include graft versus hostdisease, transplant rejection, septic shock, endotoxemia, Lymearthritis, infectious meningitis (e.g., viral, bacterial, Lymedisease-associated), an acute episode of asthma and acute episodes of anautoimmune disease.

“Inflammatory disorders” expressly include chronic inflammatorydisorders. Nonlimiting examples of chronic inflammatory disorder includeasthma, rubella arthritis, and chronic autoimmune diseases, such assystemic lupus erythematosus, psoriasis, inflammatory bowel disease,including Crohn's disease and ulcerative colitis, multiple sclerosis andrheumatoid arthritis.

The term “immune diseases” includes any immune disease, disorder orcondition caused, exasperated or mediated by IL-12 production. Suchimmune diseases may include, without limitation, rheumatoid arthritis,juvenile rheumatoid arthritis, systemic onset juvenile rheumatoidarthritis, psoriatic arthritis, ankylosing spondylitis, gastric ulcer,seronegative arthropathies, osteoarthritis, inflammatory bowel disease,ulcerative colitis, systemic lupus erythematosis, antiphospholipidsyndrome, iridocyclitis/uveitis/optic neuritis, idiopathic pulmonaryfibrosis, systemic vasculitis/wegener's granulomatosis, sarcoidosis,orchitis/vasectomy reversal procedures, allergic/atopic diseases,asthma, allergic rhinitis, eczema, allergic contact dermatitis, allergicconjunctivitis, hypersensitivity pneumonitis, transplants, organtransplant rejection, graft-versus-host disease, systemic inflammatoryresponse syndrome, sepsis syndrome, gram positive sepsis, gram negativesepsis, culture negative sepsis, fungal sepsis, neutropenic fever,urosepsis, meningococcemia, trauma/hemorrhage, burns, ionizing radiationexposure, acute pancreatitis, adult respiratory distress syndrome,rheumatoid arthritis, alcohol-induced hepatitis, chronic inflammatorypathologies, sarcoidosis, Crohn's pathology, sickle cell anemia,diabetes, nephrosis, atopic diseases, hypersensitity reactions, allergicrhinitis, hay fever, perennial rhinitis, conjunctivitis, endometriosis,asthma, urticaria, systemic anaphalaxis, dermatitis, pernicious anemia,hemolytic disease, thrombocytopenia, graft rejection of any organ ortissue, kidney transplant rejection, heart transplant rejection, livertransplant rejection, pancreas transplant rejection, lung transplantrejection, bone marrow transplant (BMT) rejection, skin allograftrejection, cartilage transplant rejection, bone graft rejection, smallbowel transplant rejection, fetal thymus implant rejection, parathyroidtransplant rejection, xenograft rejection of any organ or tissue,allograft rejection, anti-receptor hypersensitivity reactions, Gravesdisease, Raynoud's disease, type B insulin-resistant diabetes, asthma,myasthenia gravis, antibody-meditated cytotoxicity, type IIIhypersensitivity reactions, systemic lupus erythematosus, POEMS syndrome(polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy,and skin changes syndrome), polyneuropathy, organomegaly,endocrinopathy, monoclonal gammopathy, skin changes syndrome,antiphospholipid syndrome, pemphigus, scleroderma, mixed connectivetissue disease, idiopathic Addison's disease, diabetes mellitus, chronicactive hepatitis, primary biliary cirrhosis, vitiligo, vasculitis,post-MI cardiotomy syndrome, type IV hypersensitivity, contactdermatitis, hypersensitivity pneumonitis, allograft rejection,granulomas due to intracellular organisms, drug sensitivity,metabolic/idiopathic, Wilson's disease, hemachromatosis,alpha-1-antitrypsin deficiency, diabetic retinopathy, Hashimoto'sthyroiditis, osteoporosis, hypothalamic-pituitary-adrenal axisevaluation, primary biliary cirrhosis, thyroiditis, encephalomyelitis,cachexia, cystic fibrosis, neonatal chronic lung disease, chronicobstructive pulmonary disease (COPD), familial hematophagocyticlymphohistiocytosis, dermatologic conditions, psoriasis, alopecia,nephrotic syndrome, nephritis, glomerular nephritis, acute renalfailure, hemodialysis, uremia, toxicity, preeclampsia, okt3 therapy,anti-cd3 therapy, cytokine therapy, chemotherapy, radiation therapy(e.g., including but not limited to asthenia, anemia, cachexia, and thelike), chronic salicylate intoxication, and the like. See, e.g., theMerck Manual, 12th-17th Editions, Merck & Company, Rahway, N.J. (1972,1977, 1982, 1987, 1992, 1999), Pharmacotherapy Handbook, Wells et al.,eds., Second Edition, Appleton and Lange, Stamford, Conn. (1998, 2000),each entirely incorporated by reference.

The term “neurological disorder” includes any neurological disease,disorder or condition caused, exasperated or mediated by IL-12production. Such neurological disorders may include, without limitation,neurodegenerative diseases, multiple sclerosis, migraine headache, AIDSdementia complex, demyelinating diseases, such as multiple sclerosis andacute transverse myelitis; extrapyramidal and cerebellar disorders' suchas lesions of the corticospinal system; disorders of the basal gangliaor cerebellar disorders; hyperkinetic movement disorders such asHuntington's Chorea and senile chorea; drug-induced movement disorders,such as those induced by drugs which block CNS dopamine receptors;hypokinetic movement disorders, such as Parkinson's disease; Progressivesupranucleo Palsy; structural lesions of the cerebellum; spinocerebellardegenerations, such as spinal ataxia, Friedreich's ataxia, cerebellarcortical degenerations, multiple systems degenerations (Mencel,Dejerine-Thomas, Shi-Drager, and Machado-Joseph); systemic disorders(Refsum's disease, a-betalipoprotemia, ataxia, telangiectasia, andmitochondrial multi-system disorder); demyelinating core disorders, suchas multiple sclerosis, acute transverse myelitis; and disorders of themotor unit' such as neurogenic muscular atrophies (anterior horn celldegeneration, such as amyotrophic lateral sclerosis, infantile spinalmuscular atrophy and juvenile spinal muscular atrophy); Alzheimer'sdisease; Down's Syndrome in middle age; Diffuse Lewy body disease;Senile Dementia of Lewy body type; Wernicke-Korsakoff syndrome; chronicalcoholism; Creutzfeldt-Jakob disease; Subacute sclerosingpanencephalitis, Hallervorden-Spatz disease; and Dementia pugilistica,and the like. Such a method can optionally comprise administering aneffective amount of a composition or pharmaceutical compositioncomprising at least one TNF antibody or specified portion or variant toa cell, tissue, organ, animal or patient in need of such modulation,treatment or therapy. See, e.g., the Merck Manual, 16, Edition, Merck &Company, Rahway, N.J. (1992)

In the case of overlap in these definitions, the disease, condition ordisorder may be considered to be a member of any of the above listedclasses of IL-12 production-related disorders.

The term “disalt” refers to an ionic substances of formula A having acationic, diprotonated IL-12 production inhibitor compound, (Δ)(2H⁺),combined with anionic, charge balancing moieties, n(Σ). In general,disalts can be formed by

{[(Δ)(2H⁺)]²⁺·[n(Σ)]²⁻}  (A)

contacting IL-12 production inhibitor compounds ((Δ) in formula A) withBronsted acids. As used herein, the term “Bronsted acid” includes anychemical species that can be proton (H⁺) donors. While not wishing to bebound by theory, it is believed that disalt formation occurs when two ormore (H⁺)-acceptor atoms, e.g., nitrogen atoms, of the IL-12 productioninhibitor compounds are protonated by the Bronsted acid. Thus, in someembodiments, the charge balancing moieties (n(Σ) in formula A)correspond to the conjugate base of the Bronsted acid used to protonatethe IL-12 production inhibitor compounds. In other embodiments, disaltprotons ((2H⁺) in formula A) and charge balancing moieties can bereplaced in subsequent exchange reactions. For example, the disaltprotons can be exchanged, e.g., for the corresponding isotopic deuterons(2D⁺) or tritons (2T⁺), and/or the disalt charge balancing moieties canbe exchanged for other negatively charged counterions, e.g., via ionexchange chromatography methods. Disalts prepared via anion and/orcation exchange reactions of disalt starting materials are also withinthe scope of the present invention.

The disalts described herein can have a relatively high solubility,e.g., water solubility. As used herein, the “solubility” of a disalt ina solvent refers to the maximum mass of a disalt that can be dissolvedper unit volume of the solvent under accelerated dissolution conditions(50-60° C. with concommitant ultrasonic agitation of the disalt/solventtest sample). Unless otherwise indicated, solubility data is expressedin terms of units of milligrams (mg)/milliliter (mL), e.g., mg/mL.

In some embodiments, the disalt solubility in water can be at leastabout 10 mg/mL (e.g., at least about 15 mg/mL, at least about 25 mg/mL,at least about 50 mg/mL, at least about 60 mg/mL, at least about 70mg/mL, at least about 80 mg/mL, at least about 90 mg/mL, at least about100 mg/mL, at least about 150 mg/mL, at least about 200 mg/mL, at leastabout 225 mg/mL, at least about 250 mg/mL, at least about 275 mg/mL, atleast about 300 mg/mL, at least about 350 mg/mL, at least about 400mg/mL, at least about 450 mg/mL, at least about 500 mg/mL, at leastabout 550 mg/mL, at least about 600 mg/mL, at least about 650 mg/mL, atleast about 700 mg/mL, at least about 750 mg/mL, at least about 800mg/mL, at least about 850 mg/mL, at least about 900 mg/mL, at leastabout 950 mg/mL, at least about 1,000 mg/mL).

The inventors have discovered that there can be unexpected differencesin solubility between a disalt and a monosalt. As used herein, the term“monosalt” refers to ionic substances having a cationic, monoprotonatedIL-12 production inhibitor compound, e.g., (Δ)(1H⁺), combined with ananionic, charge balancing moiety, e.g., [1(Σ)]⁻¹. For example, theinventors have discovered that the solubility of a disalt can beunexpectedly higher (i.e., more positive and greater in magnitude) thanthe solubility of the corresponding monosalt (i.e., the solubility of{[(Δ)(2H⁺)]²⁺·[n(Σ)]²⁻} can be unexpectedly higher than the solubilityof {[(Δ)(1H⁺)]¹⁺·[n(Σ)]¹⁻}). Further, in some instances, the solubilityof disalt can be unexpectedly higher than monosalts differing in theidentity of either (Δ) and/or (Σ) (i.e., the solubility of{[(Δ)(2H⁺)]²⁺·[n(Σ)]²⁻} can be unexpectedly higher than the solubilityof {[(Δ′)(1H⁺)]¹⁺·[n(Σ)]¹⁻}). In some embodiments, the solubility ofdisalts can be about 2 times higher (e.g., about 3 times higher, about 4times higher, about 5 times higher, about 6 times higher, about 7 timeshigher, about 8 times higher, about 9 times higher, about 10 timeshigher, about 11 times higher, about 12 times higher, about 13 timeshigher, about 14 times higher, about 15 times higher, about 16 timeshigher, about 17 times higher, about 18 times higher, about 19 timeshigher, or about 20 times higher) than the solubility of monosalts.

Thus, disalts can have one or more advantages over monosalts as activeingredients in pharmaceutical formulations and compositions fortreatment of IL-12 related diseases or disorders, e.g., those IL-12related diseases or disorders described herein. While not wishing to bebound by theory, it is believed that, e.g., the higher solubility of thedisalts relative to the monosalts can render a disalt pharmaceuticalformulation or composition more bioavailable than a monosaltpharmaceutical formulation or composition. Again, while not wishing tobe bound by theory, it is believed that, e.g., the higher solubility ofthe disalts relative to the monosalts would increase the likelihood thatliquid-based disalt compositions and formulations could be prepared athigher concentrations than liquid-based monosalt compositions andformulations, thereby advantageously minimizing the volume of liquidcarrier needed to prepare the formulation or composition. It is alsobelieved that, e.g., the higher solubility of the disalts relative tothe monosalts would increase the likelihood that disalts could besolubilized more quickly than monosalts, thereby advantageouslyminimizing composition/formulation preparation times.

The IL-12 production inhibitor compounds used to form disalts (i.e., Δin formula A) can include compounds having any one of formulas Ia, IIa,and IIIa below (definitions for each of the variables in formulas Ia,IIa, and IIIa are as provided above).

Examples of such IL-12 production inhibitor compounds are providedbelow:

The synthesis and IL-12 production inhibitory activities of compoundshaving formulas Ia, IIa, and IIIa, e.g., compounds 1-14, as well asother useful IL-12 production inhibitor compounds, are described in,e.g., Ono, et al., U.S. Pat. No. 6,384,032, entitled “Inhibitors ofIL-12 Production;” Ono, et al., U.S. Pat. No. 6,680,315, entitled“Triazine Compounds;” Ono, et al., U.S. Pat. No. 6,693,097, entitled“Pyrimidine Compounds;” Sun, et al., U.S. Pat. No. 6,660,733, entitled“Pyrimidine Compounds;” U.S. patent application Ser. No. 10/686,505,filed on Oct. 14, 2003, entitled “Novel Compounds;” U.S. Pat. No.6,858,606, filed on Nov. 26, 2002, entitled “Pyrimidine Compounds;” U.S.patent application Ser. No. 10/985,696, filed on Nov. 10, 2004, andentitled “Pyridine Compounds;” U.S. patent application Ser. No.10/985,716, filed on Nov. 10, 2004, and entitled “Heteroaryl HydrazoneCompounds;” U.S. patent application Ser. No. 10/985,627, filed on Nov.10, 2004, and entitled “Quinazoline Compounds;” and U.S. ProvisionalPatent Application Ser. No. 60/626,609, filed Nov. 10, 2004, andentitled “Process for Trisubstituted Pyrimidine Compounds.” The entireteachings of the above mentioned patents and patent applications areincorporated herein by reference.

In some embodiments, disalt formation can occur when two or more of the(H⁺)-acceptor atoms on the IL-12 production inhibitor compounds arenitrogen atoms.

In some embodiments, it can be desirable to have R₃ in formulas Ia andIIa be an amino group; an alkylamino group; a dialkylamino group; aheterocyclyl group having at least one basic nitrogen atom, e.g., aheterocyclyl group having an alkyl or unsubstituted ring nitrogen atom,such as a morpholino group; or a heteroaryl group having at least onebasic nitrogen atom, e.g., a heteroaryl group having a nitrogen atom inwhich the lone pair does not form part of the aromatic π-electronsystem, such as a pyridinyl group.

In some embodiments, it can be desirable to have hydrazone linkages,e.g., —N(R^(c))—N═C—, and/or amino linkages, —N(R^(c))—, form part ofthe substituents R₁, L₁-B′, or R₁₂ in formulas Ia, IIa, and IIIa,respectively. R^(c) can be, independently, hydrogen or lower alkyl,e.g., CH₃, in either of the above linkages.

In some embodiments, it can be desirable to have Z be nitrogen and W beoxygen in formulas Ia and IIa and to have one of R₁₁ or R₁₂ in formulaIIIa be a morpholino group.

In some embodiments, it can be desirable to have R₃ in formulas Ia andIIa be an amino group; an alkylamino group; a dialkylamino group; aheterocyclyl group having at least one basic nitrogen atom as describedabove; or a heteroaryl group having at least one basic nitrogen atom asdescribed above; and to have either hydrazone linkages, e.g.,—N(R^(c))—N═C—, and/or amino linkages, —N(R^(c))—, form part of thesubstituents R₁ or L₁-B′ in formulas Ia and IIa, respectively.

In general, Bronsted acids useful for forming the disalts describedherein can have a pK_(a) (relative to water) in the range of betweenabout −15 and about +5 (e.g., about −15 to about −14, about −15 to about−13, about −15 to about −12, about −15 to about −11, about −15 to about−10, about −15 to about −9, about −15 to about −8, about −15 to about−7, about −15 to about −6, about −15 to about −5, about −15 to about −4,about −15 to about −3, about −15 to about −2, about −15 to about −1,about −15 to about 0, about −15 to about 0.7, about −15 to about 1,about −15 to about 2, about −15 to about 3, about −15 to about 4). Forexample, see P. Heinrich Stahl, Camille G. Wermuth. Handbook ofPharmaceutical Salts. Wiley-Vch. (2002) p. 145-149. Exemplary acidsinclude without limitation, hydrochloric (HCl); nitric (HNO₃); sulfuric(H₂SO₄); hydrobromic (HBr); hydroiodic (HI); perchloric (HClO₄);phosphoric acid (H₃PO₄); alkylsulfonic acids, e.g., methanesulfonic(CH₃SO₃H), and halogenated analogs thereof, e.g.,trifluoromethanesulfonic (CF₃SO₃H); arylsulfonic acids, e.g.,benzenesulfonic (C₆H₅SO₃H) or p-toluenesulfonic acid (p-TolSO₃H);halogenated acetic acids, e.g., trifluoroacetic (CF₃CO₂H),trichloroacetic (CCl₃CO₂H), dichloroacetic (CHCl₂CO₂H), fluoroacetic(FCH₂CO₂H), and chloroacetic (ClCH₂CO₂H)); picric ((O₂N)₃C₆H₂OH); oxalic((CO₂H)₂); citric (C(OH)(CH₂CO₂H)₂CO₂H); formic (HCO₂H); ascorbic acid;and benzoic acid (C₆H₅CO₂H) and derivatives thereof. In someembodiments, it can be desirable to use alkylsulfonic acids, e.g.,methanesulfonic (CH₃SO₃H). Other useful acids (and pKa values) aredescribed in P. Heinrich Stahl, Camille G. Wermuth. Handbook ofPharmaceutical Salts. Wiley-Vch. (2002) p. 145-149.

In some embodiments, IL-12 production inhibitor compounds can becontacted with at least two equivalents of a monoprotic Bronsted acid,e.g., HM; a diprotic Bronsted acid, e.g., H₂M; or a polyprotic Bronstedacid; e.g., H₃M, to provide disalts having formulas,{[(Δ)(2H⁺)]²⁺·[2(M⁻)]²⁻}, {[(Δ)(−2H⁺)]²⁺·[2(HM⁻)]²⁻}, and{[(Δ)(2H⁺)]²⁺·[2(H₂M⁻)]²⁻}, respectively. In some embodiments, IL-12production inhibitor compounds can be contacted with at least oneequivalent of a diprotic Bronsted acid, a polyprotic acid Bronsted, or abifunctional Bronsted acid, e.g., HM-MH, to provide disalts havingformulas, {[(Δ)(2H⁺)]²⁺·(M²⁻)]²⁻}, {[(Δ)(2H⁺)]²⁺·(HM²⁻)]²⁻}, and{[(Δ)(2H⁺)]²⁺·(⁻M-M⁻)]²⁻}, respectively.

In some embodiments, disalts can be prepared by contacting an IL-12production inhibitor compound with a solvent to form a solution orsuspension and contacting the solution or suspension with a Bronstedacid. In certain embodiments, the solution or suspension contains atmost about 1 weight percent of the IL-12 production inhibitor compound(e.g., at most about 2 weight percent of the IL-12 production inhibitorcompound, at most about 5 weight percent of the IL-12 productioninhibitor compound, at most about 10 weight percent of the IL-12production inhibitor compound, at most about 15 weight percent of theIL-12 production inhibitor compound, at most about 20 weight percent ofthe IL-12 production inhibitor compound, at most about 30 weight percentof the IL-12 production inhibitor compound, at most about 40 weightpercent of the IL-12 production inhibitor compound, or at most about 50weight percent of the IL-12 production inhibitor compound). In someembodiments, the IL-12 production inhibitor compound and the solvent canbe combined at room temperature, e.g., 25° C., to form a suspension, andthe suspension can be heated to at most about 60° C. (at most about 70°C. or at most about 80° C.) to form a solution. In other embodiments,the Bronsted acid can be added to the suspension at room temperature toform a solution. In some embodiments, it can be desirable to form theIL-12 production inhibitor compound solution, e.g., by heating, prior toadding the Bronsted acid. The Bronsted acid can be added neat, e.g., asa solid, liquid, or gas, or as a solution, e.g., HCl in ether. In someembodiments, it can be desirable to add an amount of the Bronsted acidthat is in excess of the stoichiometric amount of acid that iscalculated to be needed for disalt formation.

In some embodiments, the solvent can be a C₁-C₄ alcohol, e.g., ethanol(e.g., absolute ethanol), isopropanol, or 2,2,2-trifluoroethanol. Incertain embodiments, the solvent can be a mixed solvent systemcomprising two or more solvents and can be a homogeneous orheterogeneous solvent system. The constituent solvents of the mixedsolvent system may be present either in equal or unequal amounts, e.g.,25:75, 50:50, 90:10. Examples of mixed solvent systems include absoluteethanol/toluene and absolute ethanol/chloroform.

Disalts can be isolated, e.g., by filtration of a precipitated disalt orby evaporation of a solution containing the formed disalt. Removal ofbulk and/or residual solvents can be carried out, e.g., using one ormore of the following techniques. In some embodiments, solvent removalcan be carried out by natural evaporation (e.g., under ambientconditions with substantially no deliberate displacement of solventvapors from the vicinity of the disalt or forced evaporation). In someembodiments, solvent removal can be carried out by deliberatedisplacement of solvent vapors from the vicinity of the disalt (e.g., bya directed stream of air or an inert gas, such as nitrogen or argon).Solvent removal can be carried out in vacuo, for example, at a pressureof at least about 0.05 mm Hg (e.g., at least about 0.10 mm Hg, at leastabout 0.50 mm Hg, at least about 1 mm Hg, at least about 5 mm Hg, atleast about 10 mm Hg, at least about 30 mm Hg). In general, solventremoval can be optionally carried out, for example, at a temperature ofat most about 70° C. (e.g., at most about 60° C., at most about 50° C.,at most about 40° C., at most about 30° C., at most about 25° C.).

The extent of solvent removal can be monitored by gravimetric methods(e.g. drying of the disalt until a constant weight of the disalt isachieved) or spectroscopic techniques (e.g., removing a sample of thedisalt and obtaining a ¹H NMR spectrum of the sample to detect thesolvent).

In some embodiments, it can be desirable for the disalt to besubstantially free of solvents.

In some embodiments, it can be desirable to isolate, formulate, and/oradminister disalts that further include one or more solvents, e.g., as aresult of solvate formation or occlusion of solvent molecules into thecrystal lattice of the disalts. Disalts can have at most about 0.05weight percent (e.g., at most about 0.1 weight percent, at most about0.5 weight percent solvent, at most about 1 weight percent, at mostabout 2 weight percent, at most about 3 weight percent, at most about 4weight percent, at most about 5 weight percent, at most about 10 weightpercent, at most about 15 weight percent) of one or more solvents.

Both solvent-associated and substantially solvent-free disalts arewithin the scope of this invention.

In some instances, a disalt prepared by the methods described herein mayfurther include the corresponding monosalt. In some embodiments, it canbe desirable for the disalt to be substantially free of the monosalt.Monosalts can be removed, e.g., by recrystallization.

In some embodiments, it can be desirable to isolate, formulate, and/oradminister disalts that further include the corresponding monosalt.Disalts can have at most about 0.05 weight percent (e.g., at most about0.1 weight percent, at most about 0.5 weight percent solvent, at mostabout 1 weight percent, at most about 2 weight percent, at most about 3weight percent, at most about 4 weight percent, at most about 5 weightpercent, at most about 10 weight percent, at most about 15 weightpercent, at most about 20 weight percent, at most about 25 weightpercent, at most about 30 weight percent, at most about 35 weightpercent, at most about 40 weight percent, at most about 45 weightpercent,) of the monosalt.

Both monosalt-associated and substantially monosalt-free disalts arewithin the scope of this invention.

Also within the scope of this invention is a pharmaceutical compositionthat contains an effective amount of one or more of the disalts of thisinvention and a pharmaceutically acceptable carrier. Further, thepresent invention covers a method of administering an effective amountof such a disalt to a subject in need of treatment of IL-12 productionrelated diseases (e.g., rheumatoid arthritis, sepsis, Crohn's disease,multiple sclerosis, psoriasis, or insulin-dependent diabetes mellitus).“An effective amount” refers to the amount of the disalt which isrequired to confer a therapeutic effect on the treated subject. Theinterrelationship of dosages for animals and humans (based on milligramsper meter squared of body surface) is described in Freireich et al.,(1966) Cancer Chemother Rep 50: 219. Body surface area may beapproximately determined from height and weight of the patient. See,e.g., Scientific Tables, Geigy Pharmaceuticals, Ardley, N.Y., 1970, 537.An effective amount of the disalt of this invention can range from about0.001 mg/Kg to about 1000 mg/Kg. Effective doses will also vary, asrecognized by those skilled in the art, depending on the diseasestreated, route of administration, excipient usage, and the possibilityof co-usage with other therapeutic treatments such as use of otheragents.

To practice the method of the present invention, a disalt, as acomponent of a pharmaceutical composition, can be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

A sterile injectable composition, for example, a sterile injectableaqueous or oleaginous suspension, can be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation can also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that can be employed are mannitol, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils are conventionally employed as a solvent or suspending medium(e.g., synthetic mono- or diglycerides). Fatty acids, such as oleic acidand its glyceride derivatives are useful in the preparation ofinjectables, as are natural pharmaceutically-acceptable oils, such asolive oil or castor oil, especially in their polyoxyethylated versions.These oil solutions or suspensions can also contain a long-chain alcoholdiluent or dispersant, or carboxymethyl cellulose or similar dispersingagents. Other commonly used surfactants such as Tweens or Spans or othersimilar emulsifying agents or bioavailability enhancers which arecommonly used in the manufacture of pharmaceutically acceptable solid,liquid, or other dosage forms can also be used for the purposes offormulation.

A composition for oral administration can be any orally acceptabledosage form including, but not limited to, capsules, tablets, emulsionsand aqueous suspensions, dispersions and solutions. In the case oftablets for oral use, carriers which are commonly used include lactoseand corn starch. Lubricating agents, such as magnesium stearate, arealso typically added. For oral administration in a capsule form, usefuldiluents include lactose and dried corn starch. When aqueous suspensionsor emulsions are administered orally, the active ingredient can besuspended or dissolved in an oily phase combined with emulsifying orsuspending agents. If desired, certain sweetening, flavoring, orcoloring agents can be added. A nasal aerosol or inhalation compositioncan be prepared according to techniques well-known in the art ofpharmaceutical formulation and can be prepared as solutions in saline,employing benzyl alcohol or other suitable preservatives, absorptionpromoters to enhance bioavailability, fluorocarbons, and/or othersolubilizing or dispersing agents known in the art. A disalt of thisinvention can also be administered in the form of suppositories forrectal administration.

The carrier in the pharmaceutical composition must be “acceptable” inthe sense of being compatible with the active ingredient of theformulation (and preferably, capable of stabilizing it) and notdeleterious to the subject to be treated. For example, solubilizingagents such as cyclodextrins, which form specific, more solublecomplexes with the disalts of this invention, or one or moresolubilizing agents, can be utilized as pharmaceutical excipients fordelivery of the pyridine compounds. Examples of other carriers includecolloidal silicon dioxide, magnesium stearate, cellulose, sodium laurylsulfate, and D&C Yellow #10.

In certain embodiments, pharmaceutical compositions and dosage forms ofthe invention comprise one or more active ingredients in relativeamounts and formulated in such a way that a given pharmaceuticalcomposition or dosage form inhibits the uptake of calcium. Preferredpharmaceutical compositions and dosage forms comprise a disalt describedherein, or a pharmaceutically acceptable prodrug, solvate, or clathratethereof, optionally in combination with one or more additional activeagents.

The methods for treating or preventing disorders associated withexcessive bone loss in a patient in need thereof can further compriseadministering to the patient being administered a compound of thisinvention, an effective amount of one or more other therapeutic agents.Such therapeutic agents may include other therapeutic agents such asthose conventionally used to prevent or treat disorders associated withexcessive bone resorption or symptoms thereof. For example, such otheragents include anti-resorptive agents for example progestins,polyphosphonates, bisphosphonate(s), estrogen agonists/antagonists,estrogen (such as Premarin®), estrogen/progestin combinations, andestrogen derivatives (such as estrone, estriol or 17α,17β-ethynylestradiol).

In such combination therapy treatment, both the compounds of thisinvention and the other drug agent(s) are administered to mammals (e.g.,humans, male or female) by conventional methods. The agents may beadministered in a single dosage form or in separate dosage forms.Effective amounts of the other therapeutic agents are well known tothose skilled in the art. However, it is well within the skilledartisan's purview to determine the other therapeutic agent's optimaleffective-amount range. In one embodiment of the invention where anothertherapeutic agent is administered to an animal, the effective amount ofthe compound of this invention is less than its effective amount wouldbe where the other therapeutic agent is not administered. In anotherembodiment, the effective amount of the conventional agent is less thanits effective amount would be where the compound of this invention isnot administered. In this way, undesired side effects associated withhigh doses of either agent may be minimized. Other potential advantages(including without limitation improved dosing regimens and/or reduceddrug cost) will be apparent to those of skill in the art.

Exemplary progestins are available from commercial sources and include:algestone acetophenide, altrenogest, amadinone acetate, anagestoneacetate, chlormadinone acetate, cingestol, clogestone acetate,clomegestone acetate, delmadinone acetate, desogestrel, dimethisterone,dydrogesterone, ethynerone, dthynodiol diacetate, etonogestrel,flurogestone acetate, gestaclone, gestodene, gestonorone caproate,gestrinone, haloprogesterone, hydroxyprogesterone, caproate,levonorgestrel, lynestrenol, medrogestone, medroxyprogesterone acetate,melengestrol acetate, methynodiol diacetate, norethindrone,norethindrone acetate, norethynodrel, norgestimate, norgestomet,norgestrel, oxogestone phenpropionate, progesterone, quingestanolacetate, quingestrone, and tigestol. Preferred progestins aremedroxyprogestrone, norethindrone and norethynodrel.

Exemplary bone resorption inhibiting polyphosphonates includepolyphosphonates of the type disclosed in U.S. Pat. No. 3,683,080.Preferred polyphosphonates are geminal dipolyphosphonates (also referredto as bis-phosphonates). Tiludronate disodium is an especially preferredpolyphosphonate. Ibandronic acid is an especially preferredpolyphosphonate. Alendronate is an especially preferred polyphosphonate.Zoledronic acid is an especially preferred polyphosphonate. Otherpreferred polyphosphonates are6-amino-1-hydroxy-hexylidene-bisphosphonic acid and1-hydroxy-3-(methylpentylamino)-propylidene-bisphosphonic acid. Thepolyphosphonates may be administered in the form of the acid, or of asoluble alkali metal salt or alkaline earth metal salt. Hydrolyzableesters of the polyphosphonates are likewise included. Specific examplesinclude ethane-1-hydroxy-1,1-diphosphonic acid, methane diphosphonicacid, pentane-1-hydroxy-1,1-diphosphonic acid, methane dichlorodiphosphonic acid, methane hydroxy diphosphonic acid,ethane-1-amino-1,1-diphosphonic acid, ethane-2-amino-1,1-diphosphonicacid, propane-3-amino-1-hydroxy-1,1-diphosphonic acid,propane-N,N-dimethyl-3-amino-1-hydroxy-1,1-diphosphonic acid,propane-3,3-dimethyl-3-amino-1-hydroxy-1,1-diphosphonic acid, phenylamino methane diphosphonic acid, N,N-dimethylamino methane diphosphonicacid, N(2-hydroxyethyl)amino methane diphosphonic acid,butane-4-amino-1-hydroxy-1,1-diphosphonic acid,pentane-5-amino-1-hydroxy-1,1-diphosphonic acid,hexane-6-amino-1-hydroxy-1,1-diphosphonic acid and pharmaceuticallyacceptable esters and salts thereof.

In particular, the disalts of this invention may be combined with amammalian estrogen agonist/antagonist. Any estrogen agonist/antagonistmay be used for this purpose. The term estrogen agonist/antagonistrefers to compounds which bind with the estrogen receptor, inhibit boneturnover and/or prevent bone loss. In particular, estrogen agonists areherein defined as chemical compounds capable of binding to the estrogenreceptor sites in mammalian tissue, and mimicking the actions ofestrogen in one or more tissue. Estrogen antagonists are herein definedas chemical compounds capable of binding to the estrogen receptor sitesin mammalian tissue; and blocking the actions of estrogen in one or moretissues. Such activities are readily determined by those skilled in theart of standard assays including estrogen receptor binding assays,standard bone histomorphometric and densitometer methods, and E. FEriksen et al., Bone Histomorphometry, Raven Press, New York, pp. 1-74(1994); S. J. Grier et. al., The Use of Dual-Energy X-Ray AbsorptiometryIn Animals, Inv. Radiol. 31(1): 50-62 (1996); Wahner H. W. and FogelmanI., The Evaluation of Osteoporosis: Dual Energy X-Ray Absorptiometry inClinical Practice., Martin Dunitz Ltd., London, pp. 1-296 (1994)). Avariety of these compounds are described and referenced below.

A preferred estrogen agonist/antagonist is droloxifene: (phenol,3-(1-(4-(2-(dimethylamino)ethoxy)phenyl)-2-phenyl-1-butenyl)-, (E)-) andrelated compounds which are disclosed in U.S. Pat. No. 5,047,431.Another preferred estrogen agonist/antagonist is3-(4-(1,2-diphenyl-but-1-enyl)-phenyl)-acrylic acid, which is disclosedin Wilson et al., Endocrinology 138: 3901-11 (1997). Another preferredestrogen agonist/antagonist is tamoxifen:(ethanamine,2-(-4-(1,2-diphenyl-1-butenyl)phenoxy)-N,N-dimethyl,(Z)-2-,2-hydroxy-1,2,3-propanetricarboxylate(1:1)) and related compoundswhich are disclosed in U.S. Pat. No. 4,536,516. Another related compoundis 4-hydroxy tamoxifen which is disclosed in U.S. Pat. No. 4,623,660.

A preferred estrogen agonist/antagonist is raloxifene: (methanone,(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thien-3-yl)(4-(2-(1-piperidinyl)ethoxy)phenyl)hydrochloride)which is disclosed in U.S. Pat. No. 4,418,068. Another preferredestrogen agonist/antagonist is toremifene: (ethanamine,2-(4-(4-chloro-1,2-diphenyl-1-butenyl)phenoxy)-N,N-dimethyl-,(Z)-,2-hydroxy-1,2,3-propanetricarboxylate (1:1) which is disclosed inU.S. Pat. No. 4,996,225. Another preferred estrogen agonist/antagonistis centchroman:1-(2-((4-methoxy-2,2,dimethyl-3-phenyl-chroman-4-yl)-phenoxy)-ethyl)-pyrrolidine,which is disclosed in U.S. Pat. No. 3,822,287. Also preferred islevormeloxifene. Another preferred estrogen agonist/antagonist isidoxifene:(E)-1-(2-(4-(1-(4-iodo-phenyl)-2-phenyl-but-1-enyl)-phenoxy)-ethyl)-pyrrolidinone,which is disclosed in U.S. Pat. No. 4,839,155. Another preferredestrogen agonist/antagonist is2-(4-methoxy-phenyl)-3-[4-(2-piperidin-1-yl-ethoxy)-phenoxy]-benzo[b]thiophen-6-olwhich is disclosed in U.S. Pat. No. 5,488,058. Another preferredestrogen agonist/antagonist is6-(4-hydroxy-phenyl)-5-(4-(2-piperidin-1-yl-ethoxy)-benzyl)-naphthalen-2-olwhich is disclosed in U.S. Pat. No. 5,484,795. Another preferredestrogen agonist/antagonist is(4-(2-(2-aza-bicyclo[2.2.1]hept-2-yl)-ethoxy)-phenyl)-(6-hydroxy-2-(4-hydroxy-phenyl)-benzo[b]thiophen-3-yl)-methanonewhich is disclosed, along with methods of preparation, in PCTpublication no. WO 95/10513 assigned to Pfizer Inc. Other preferredestrogen agonist/antagonists include compounds as described in U.S. Pat.No. 5,552,412. Especially preferred compounds described therein are:cis-6-(4-fluoro-phenyl)-5-(4-(2-piperidin-1-yl-ethoxy)-phenyl)-5,6,7,8-tetrahydro-naphthalene-2-ol;(−)-cis-6-phenyl-5-(4-(2-pyrrolidin-1-yl-ethoxy)-phenyl)-5,6,7,8-tetrahydro-naphthalene-2-ol;cis-6-phenyl-5-(4-(2-pyrrolidin-1-yl-ethoxy)-phenyl)-5,6,7,8-tetrahydro-naphthalene-2-ol;cis-1-(6′-pyrrolodinoethoxy-3′-pyridyl)-2-phenyl-6-hydroxy-1,2,3,4-tetrahydronaphthalene;1-(4′-pyrrolidinoethoxyphenyl)-2-(4″-fluorophenyl)-6-hydroxy-1,2,3,4-tetrahydroisoquinoline;cis-6-(4-hydroxyphenyl)-5-(4-(2-piperidin-1-yl-ethoxy)-phenyl)-5,6,7,8-tetrahydro-naphthalene-2-ol;and1-(4′-pyrrolidinolethoxyphenyl)-2-phenyl-6-hydroxy-1,2,3,4-tetrahydroisoquinoline.Other estrogen agonist/antagonists are described in U.S. Pat. No.4,133,814. U.S. Pat. No. 4,133,814 discloses derivatives of2-phenyl-3-aroyl-benzothiophene and2-phenyl-3-aroylbenzothiophene-1-oxide.

Those skilled in the art will recognize that other bone anabolic agents,also referred to as bone mass augmenting agents, may be used inconjunction with the compounds of this invention. A bone mass augmentingagent is a compound that augments bone mass to a level which is abovethe bone fracture threshold as detailed in the World Health OrganizationStudy World Health Organization, “Assessment of Fracture Risk and itsApplication to Screening for Postmenopausal Osteoporosis (1994). Reportof a WHO Study Group. World Health Organization Technical Series 843.”Any prostaglandin, or prostaglandin agonist/antagonist may be used incombination with the compounds of this invention. Those skilled in theart will recognize that IGF-1, sodium fluoride, parathyroid hormone(PTH), active fragments of parathyroid hormone, growth hormone or growthhormone secretagogues may also be used. The following paragraphsdescribes in greater detail exemplary compounds that may be administeredin combination with compounds of this invention

Prostaglandins: The term prostaglandin refers to compounds which areanalogs of the natural prostaglandins PGD₁, PGD₂, PGE₂, PGE₁ and PGF₂which are useful in the treatment of osteoporosis and other disordersassociated with excessive osteoclastic bone resorption. These compoundsbind to the prostaglandins receptors. Such binding is readily determinedby those skilled in the art of standard assays (e.g., S. An et al.,Cloning and Expression of the EP₂ Subtype of Human Receptors forProstaglandin E₂ Biochemical and Biophysical Research Communications,197(1): 263-270 (1993)).

Prostaglandins are alicyclic compounds related to the basic compoundprostanoic acid. The carbon atoms of the basic prostaglandin arenumbered sequentially from the carboxylic carbon atom through thecyclopentyl ring to the terminal carbon atom on the adjacent side chain.Normally the adjacent side chains are in the trans orientation. Thepresence of an oxo group at C-9 of the cyclopentyl moiety is indicativeof a prostaglandin within the E class while PGE₂ contains a transunsaturated double bond at the C₁₃-C₁₄ and a cis double bond at theC₅-C₆ position.

A variety of prostaglandins are described and referenced below. However,other prostaglandins will be known to those skilled in the art.Exemplary prostaglandins are disclosed in U.S. Pat. Nos. 4,171,331 and3,927,197. Norrdin et al., The Role of Prostaglandins in Bone in Vivo,Prostaglandins Leukotriene Essential Fatty Acids 41: 139-150 (1990) is areview of bone anabolic prostaglandins. Any prostaglandinagonist/antagonist may be used in combination with the compounds of thisinvention. The term prostaglandin agonist/antagonist refers to compoundswhich bind to prostaglandin receptors (e.g., An S. et al., Cloning andExpression of the EP₂ Subtype of Human Receptors for Prostaglandin E₂,Biochemical and Biophysical Research Communications 197(1): 263-70(1993)) and mimic the action of prostaglandin in vivo (e.g., stimulatebone formation and increase bone mass). Such actions are readilydetermined by those skilled in the art of standard assays. Eriksen E. F.et al., Bone Histomorphometry, Raven Press, New York, 1994, pp. 1-74; S.J. Grier et al., The Use of Dual-Energy X-Ray Absorptiometry In Animals,Inv. Radiol. 31(1): 50-62 (1996); H. W. Wahner and I. Fogelman, TheEvaluation of Osteoporosis: Dual Energy X-Ray Absorptiometry in ClinicalPractice, Martin Dunitz Ltd. London, pp. 1-296 (1994). A number of thesecompounds are described and reference below. However, otherprostaglandin agonists/antagonists will be known to those skilled in theart. Exemplary prostaglandin agonists/antagonists are disclosed asfollows. U.S. Pat. No. 3,932,389 discloses2-descarboxy-2-(tetrazol-5-yl)-11-desoxy-15-substituted-omega-pentanorprostaglandinsuseful for bone formation activity. U.S. Pat. No. 4,018,892, discloses16-aryl-13,14-dihydro-PGE₂ p-biphenyl esters useful for bone formationactivity. U.S. Pat. No. 4,219,483, discloses 2,3,6-substituted-4-pyronesuseful for bone formation activity. U.S. Pat. No. 4,132,847, discloses2,3,6-substituted-4-pyrones useful for bone formation activity. U.S.Pat. No. 4,000,309, discloses 16-aryl-13,14-dihydro-PGE₂ p-biphenylesters useful for bone formation activity. U.S. Pat. No. 3,982,016,discloses 16-aryl-13,14-dihydro-PGE₂ p-biphenyl esters useful for boneformation activity. U.S. Pat. No. 4,621,100, discloses substitutedcyclopentanes useful for bone formation activity. U.S. Pat. No.5,216,183, discloses cyclopentanones useful for bone formation activity.

Sodium fluoride may be used in combination with the compounds of thisinvention. The term sodium fluoride refers to sodium fluoride in all itsforms (e.g., slow release sodium fluoride, sustained release sodiumfluoride). Sustained release sodium fluoride is disclosed in U.S. Pat.No. 4,904,478. The activity of sodium fluoride is readily determined bythose skilled in the art of biological protocols.

Bone morphogenetic protein may be used in combination with the disaltsof this invention (e.g., see Ono et al., Promotion of the OsteogeneticActivity of Recombinant Human Bone Morphogenetic Protein byProstaglandin E₁, Bone 19(6): 581-588 (1996)).

Any parathyroid hormone (PTH) may be used in combination with thecompound of this invention. The term parathyroid hormone refers toparathyroid hormone, fragments or metabolites thereof and structuralanalogs thereof which can stimulate bone formation and increase bonemass. Also included are parathyroid hormone related peptides and activefragments and analogs of parathyroid related peptides (see PCTpublication No. WO 94/01460). Such bone anabolic functional activity isreadily determined by those skilled in the art of standard assays. Avariety of these compounds are described and referenced below. However,other parathyroid hormone will be known to those skilled in the art.Exemplary parathyroid hormones are disclosed in the followingreferences. “Human Parathyroid Peptide Treatment of VertebralOsteoporosis”, Osteoporosis Int., 3, (Supp 1): 199-203. “PTH 1-34Treatment of Osteoporosis with Added Hormone Replacement Therapy:Biochemical, Kinetic and Histological Responses” Osteoporosis Int. 1:162-170.

Any growth hormone or growth hormone secretagogue may be used incombination with the compounds of this invention. The term growthhormone secretagogue refers to a compound which stimulates the releaseof growth hormone or mimics the action of growth hormone (e.g.,increases bone formation leading to increased bone mass). Such actionsare readily determined by those skilled in the art of standard assayswell known to those of skill in the art. A variety of these compoundsare disclosed in the following published PCT patent applications: WO95/14666; WO 95/13069; WO 94/19367; WO 94/13696; and WO 95/34311.However, other growth hormones or growth hormone secretagogues will beknown to those skilled in the art. In particular, a preferred growthhormone secretagogue isN-[1(R)-[1,2-Dihydro-1-methanesulfonylspiro[3H-indole-3,4′-piperidin]-1′-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-amino-2-methylpropanamide:MK-667.Other preferred growth hormone secretagogues include2-amino-N-(2-(3a-(R)-benzyl-2-methyl-3-oxo-2,3,3a,4,6,7-hexahydro-pyrazolo-[4,3-c]pyridin-5-yl)-1-(R)-benzyloxymethyl-2-oxo-ethyl)-isobutyramideor its L-tartaric acid salt;2-amino-N-(1-(R)-benzyloxymethyl-2-(3a-(R)-(4-fluoro-benzyl)-2-methyl-3-oxo-2,3,3a,4,6,7-hexahydro-pyrazolo[4,3-c]pyridin-5-yl)-2-oxo-ethyl)isobutyramide;2-amino-N-(2-(3a-(R)-benzyl-3-oxo-2,3,3a,4,6,7-hexahydro-pyrazolo[4,3-c]pyridin-5-yl)-1-(R)benzyloxymethyl-2-oxo-ethyl)isobutyramide;and2-amino-N-(1-(2,4-difluoro-benzyloxymethyl)-2-oxo-2-(3-oxo-3a-pyridin-2-ylmethyl-2-(2,2,2-trifluoro-ethyl)-2,3,3a,4,6,7-hexahydro-pyrazolo[4,3-c]pyridin-5-yl)-ethyl)-2-methyl-propionamide.

The other therapeutic agent can be a steroid or a non-steroidalanti-inflammatory agent. Useful non-steroidal anti-inflammatory agents,include, but are not limited to, aspirin, ibuprofen, diclofenac,naproxen, benoxaprofen, flurbiprofen, fenoprofen, flubufen, ketoprofen,indoprofen, piroprofen, carprofen, oxaprozin, pramoprofen, muroprofen,trioxaprofen, suprofen, aminoprofen, tiaprofenic acid, fluprofen,bucloxic acid, indomethacin, sulindac, tolmetin, zomepirac, tiopinac,zidometacin, acemetacin, fentiazac, clidanac, oxpinac, mefenamic acid,meclofenamic acid, flufenamic acid, niflumic acid, tolfenamic acid,diflurisal, flufenisal, piroxicam, sudoxicam, isoxicam; salicylic acidderivatives, including aspirin, sodium salicylate, choline magnesiumtrisalicylate, salsalate, diflunisal, salicylsalicylic acid,sulfasalazine, and olsalazin; para-aminophennol derivatives includingacetaminophen and phenacetin; indole and indene acetic acids, includingindomethacin, sulindac, and etodolac; heteroaryl acetic acids, includingtolmetin, diclofenac, and ketorolac; anthranilic acids (fenamates),including mefenamic acid, and meclofenamic acid; enolic acids, includingoxicams (piroxicam, tenoxicam), and pyrazolidinediones (phenylbutazone,oxyphenthartazone); and alkanones, including nabumetone andpharmaceutically acceptable salts thereof and mixtures thereof. For amore detailed description of the NSAIDs, see Paul A. Insel,Analgesic-Antipyretic and Antiinflammatory Agents and Drugs Employed inthe Treatment of Gout, in Goodman & Gilman's The Pharmacological Basisof Therapeutics 617-57 (Perry B. Molinhoff and Raymond W. Ruddon eds.,9^(th) ed 1996) and Glen R. Hanson, Analgesic, Antipyretic andAnti-Inflammatory Drugs in Remington: The Science and Practice ofPharmacy Vol II 1196-1221 (A. R. Gennaro ed. 19th ed. 1995) which arehereby incorporated by reference in their entireties.

For arthritis, inflammation-mediated bone loss and other disorders thathave an inflammatory component, preferred conventional treatments foruse in combination therapy with the compounds and compositions of thisinvention include (without limitation) naproxen sodium (Anaprox® andAnaprox® DS, Roche), flurbiprofen (Ansaid®; Pharmacia), diclofenacsodium+misoprostil (Arthrotec®, Searle), valdecoxib (Bextra®,Pharmacia), diclofenac potassium (Cataflam® and Voltaren®, Novartis),celecoxib (Celebrex®, Pfizer), sulindac (Clinoril®, Merck), oxaprozin(Daypro®, Pharmacia), salsalate (Disalcid®, 3M), diflunisal (Dolobid®,Merck), naproxen sodium (EC Naprosyn®, Roche), piroxicam (Feldene®,Pfizer), indomethacin (Indocin® and Indocin SR®, Merck), etodolac(Lodine® and Lodine XL®, Wyeth), meloxicam (Mobic®, BoehringerIngelheim), ibuprofen (Motrin®, Pharmacia), naproxen (Naprelan®, Elan),naproxen (Naprosyn®, Roche), ketoprofen (Orudis® and Oruvail®, Wyeth),nabumetone (Relafen®, SmithKline), tolmetin sodium (Tolectin®, McNeil),choline magnesium trisalicylate (Trilisate®, Purdue Fredrick), androfecoxib (Vioxx®, Merck).

In any case where pain in a component of the target disorder, the othertherapeutic agent can be an analgesic. Useful analgesics include, butare not limited to, phenacetin, butacetin, acetaminophen, nefopam,acetoamidoquinone, and mixtures thereof.

For use against osteoporosis, Paget's disease and other disordersassociated with bone deterioration, preferred conventional agents thatmay be used in combination with compounds and compositions of thisinvention include (without limitation) bisphosphonates (such asetidronate (Didronel®, Procter & Gamble), pamidronate (Aredia®,Novartis), and alendronate (Fosamax®, Merck)), tiludronate (Skelid®,Sanofi-Synthelabo, Inc.), risedronate (Actonel®, Procter &Gamble/Aventis), calcitonin (Miacalcin®), estrogens (Climara®, Estrace®,Estraderm®, Estratab®, Ogen®, Ortho-Est®, Vivelle®, Premarin®, andothers) estrogens and progestins (Activella™, FemHrt®, Premphase®,Prempro®, and others), parathyroid hormone and portions thereof, such asteriparatide (Forteo®, Eli Lilly and Co.), selective estrogen receptormodulators (SERMs) (such as raloxifene (Evista®)) and treatmentscurrently under investigation (such as other parathyroid hormones,sodium fluoride, vitamin D metabolites, and other bisphosphonates andselective estrogen receptor modulators).

Any method of the present invention can comprise administering aneffective amount of a composition or pharmaceutical compositioncomprising at least one disalt of this invention to a cell, tissue,organ, animal or patient in need of such modulation, treatment ortherapy. Such a method can optionally further comprise co-administrationor combination therapy for treating an IL-12 production relateddisorder, wherein the administering further comprises administeringbefore, concurrently with, and/or after the compound of this invention,at least one additional active agent selected from a TNF antagonist(e.g., but not limited to a TNF antibody or fragment, a soluble TNFreceptor or fragment, fusion proteins thereof, or a small molecule TNFantagonist), an antirheumatic (e.g., methotrexate, auranofin,aurothioglucose, azathioprine, etanercept, gold sodium thiomalate,hydroxychloroquine sulfate, leflunomide, sulfasalzine), a musclerelaxant, a narcotic, a non-steroid anti-inflammatory drug (NSAID), ananalgesic, an anesthetic, a sedative, a local anesthetic, aneuromuscular blocker, an antimicrobial (e.g., aminoglycoside, anantifungal, an antiparasitic, an antiviral, a carbapenem, cephalosporin,a fluoroquinolone, a macrolide, a penicillin, a sulfonamide, atetracycline, another antimicrobial), an antipsoriatic, acorticosteroid, an anabolic steroid, a diabetes related agent, amineral, a nutritional, a thyroid agent, a vitamin, a calcium relatedhormone, an antidiarrheal, an antitussive, an antiemetic, an antiulcer,a laxative, an anticoagulant, an erythropoietin (e.g., epoetin alpha), afilgrastim (e.g., G-CSF, Neupogen), a sargramostim (GM-CSF, Leukine), animmunization, an immunoglobulin, an immunosuppressive (e.g.,basiliximab, cyclosporine, daclizumab), a growth hormone, a hormonereplacement drug, an estrogen receptor modulator, a mydriatic, acycloplegic, an alkylating agent, an antimetabolite, a mitoticinhibitor, a radiopharmaceutical, an antidepressant, antimanic agent, anantipsychotic, an anxiolytic, a hypnotic, a sympathomimetic, astimulant, donepezil, tacrine, an asthma medication, a beta agonist, aninhaled steroid, a leukotriene inhibitor, a methylxanthine, a cromolyn,an epinephrine or analog, domase alpha (Pulmozyme), a cytokine or acytokine antagonist. Suitable dosages are well known in the art. See,e.g., Wells et al., eds., Pharmacotherapy Handbook, 2nd Edition,Appleton and Lange, Stamford, Conn. (2000); PDR Pharmacopoeia, TarasconPocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, LomaLinda, Calif. (2000), each of which references are entirely incorporatedherein by reference.

TNF antagonists suitable for compositions, combination therapy,co-administration, devices and/or methods of the present inventioninclude, but are not limited to, anti-TNF antibodies (such as, Remicade(Infliximab) or Humira (adalimumab)) for example, or , antigen-bindingfragments thereof, and receptor molecules which bind specifically to TNF(such as, for example, Enbrel (Etanercept)); compounds which preventand/or inhibit TNF synthesis, TNF release or its action on target cells,such as thalidomide, tenidap, phosphodiesterase inhibitors (e.g.,pentoxifylline and rolipram), A2b adenosine receptor agonists and A2badenosine receptor enhancers; compounds which prevent and/or inhibit TNFreceptor signalling, such as mitogen activated protein (MAP) kinaseinhibitors; compounds which block and/or inhibit membrane TNF cleavage,such as metalloproteinase inhibitors; compounds which block and/orinhibit TNF activity, such as angiotensin converting enzyme (ACE)inhibitors (e.g., captopril); and compounds which block and/or inhibitTNF production and/or synthesis, such as MAP kinase inhibitors.

For clarification, a “tumor necrosis factor antibody,” “TNF antibody,”“TNF antibody,” or fragment and the like decreases, blocks, inhibits,abrogates or interferes with TNF activity in vitro, in situ and/orpreferably in vivo. For example, a suitable TNF human antibody of thepresent invention can bind TNF-α and includes anti-TNF antibodies,antigen-binding fragments thereof, and specified mutants or domainsthereof that bind specifically to TNFa. A suitable TNF antibody orfragment can also decrease block, abrogate, interfere, prevent and/orinhibit TNF RNA, DNA or protein synthesis, TNF release, TNF receptorsignaling, membrane TNF cleavage, TNF activity, TNF production and/orsynthesis.

The foregoing and other useful combination therapies will be understoodand appreciated by those of skill in the art. Potential advantages ofsuch combination therapies include the ability to use less of each ofthe individual active ingredients to minimize toxic side effects,synergistic improvements in efficacy, improved ease of administration oruse and/or reduced overall expense of compound preparation orformulation.

The biological activities of a disalt can be evaluated by a number ofcell-based assays. One of such assays can be conducted using cells fromhuman peripheral blood mononuclear cells (PBMC) or human monocytic cellline (THP-1). The cells are stimulated with a combination of humaninterferon-γ (IFNγ) and lipopolysaccharide or a combination of IFNγ andStaphylococcus aureus Cowan I in the presence of a test compound. Thelevel of inhibition of IL-12 production can be measured by determiningthe amount of p70 by using a sandwich ELISA assay with anti-human IL-12antibodies. IC₅₀ of the test compound can then be determined.Specifically, PBMC or THP-1 cells are incubated with the test compound.Cell viability was assessed using the bioreduction of MTS[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium](Promega, Madison, Wis.).

A disalt can also be evaluated by animal studies. For example, one ofsuch studies involves the ability of a test compound to treat adjuvantarthritis (i.e., a IL-12 production related disorder) in rats.

Responsiveness of a particular condition, disease or disorder to disaltsand compositions of this invention can be measured directly bycomparison against conventional drugs, or can be inferred based on anunderstanding of disease etiology and progression. There are a number ofcellular and bone resorption assay systems that are widely accepted inthe art as predictive of in vivo effects. As the bone resorption assayuses material that includes all bone cells, it is an ex vivo assay.Thus, the showing that a disalt of this invention inhibits boneresorption in these assays is evidence of the clinical utility of thesefor treating or preventing conditions associated with excessive boneloss. Various scientific publications (such as Carano et al. J. Clin.Invest. 85: 456-461 (1990); Blair & Schlesinger, The Biology andPhysiology of the Osteoclast, CRC Press, Eds., Gay, C. V. and Rifkin, B.R., pp. 259-288 (1992); and Vaananen et al., J. Cell Biology 111:1305-1311 (1990)) support the fact that such assays are accepted asbeing predictive of in vivo activity. Furthermore, the in vitro effectsof Herbimycin A on bone resorption were shown to correlate with in vivoactivity (Yoneda et al., J. Clin. Invest. 91: 2791-95 (1993)).

Without further elaboration, it is believed that the above descriptionhas adequately enabled the present invention. The following specificembodiments are, therefore, to be construed as merely illustrative, andnot limitative of the remainder of the disclosure in any way whatsoever.All of the publications cited herein are hereby incorporated byreference in their entirety.

Examples Example 1 General Procedures for Disalt Formation

Procedure I. A stirred suspension of the IL-12 production inhibitorcompound in absolute ethanol (in amount sufficient to obtain 0.33-0.34 Msolution) is heated to 70° C. until all solids are dissolved. To thestirred, heated (solution was allowed to cool down to room temperaturewhen hydrogen chloride solution in ether is used) solution anappropriate acid or solution thereof, 2 equivalents, is addedportionwise (dropwise if liquid or solution and in 4-5 portions ifsolid), heating (65-70° C.). The solution is stirred for an additional 2minutes, and then is allowed to cool to room temperature and is allowedto stand during precipitation of the disalt at room temperature for 2hours or until precipitation of the disalt was complete. In instanceswhen precipitation is slow or difficult, stirring often facilitatesprecipitation. The disalt is filtered, washed with two portions ofanhydrous ethanol:ethyl ether 1:2 mixture and then with anhydrous ethylether, and is immediately (to minimize the possibility of solventocclusion) vacuum-dried (ca. 2 mm Hg) at 50-60° C. (water bath) forabout 45 minutes to 2 hours (drying is continued until the disaltachieves a constant weight and/or no ethanol is detected by ¹H NMR). Thedisalt is then dried in vacuo further over a desiccant for an additional2 hours at ca. 2 mm Hg over DRIERITE or phosphorous pentoxide. Thedisalts are generally colorless solids and are stored in thelight-protected bottles.

Procedure II. A stirred suspension of the IL-12 production inhibitorcompound in a solvent or mixed solvent system (absolute ethanol, mixtureof absolute ethanol and toluene (1:1), or mixture of absolute ethanoland chloroform (1:1) is heated to 70° C. until all solids are dissolved.To the stirred, hot solution is added dropwise 2 equivalents of methanesulfonic acid. Heating (65-70° C.) and stiffing is continued for 2 min,and the resultant solution is left at room temperature for precipitation(in those instances in which chloroform is used, the chloroform wasremoved under reduced pressure and equal amount of toluene was added toachieve precipitation). The disalt is filtered, washed with two portionsof anhydrous ethanol:ether (1:2) then with anhydrous ether, andimmediately vacuum-dried at 50-60° C. (water bath) for 2 hours to affordcorresponding dimesylates. The disalt is then dried in vacuo furtherover a desiccant for an additional 2 hours at ca. 2 mmHg over DRIERITEor phosphorous pentoxide. The disalts are generally colorless solids andare stored in the light-protected bottles.

Procedure III. Alternatively, an ethanol suspension of the IL-12production inhibitor compound is treated with 2 equivalents of methanesulfonic acid and stirred at room temperature until all solidsdissolved. Disalt formation is then carried out as described above.

Example 2N-(1H-indol-3-ylmethylene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazine(2) dimethanesulfonate (dimesylate) {[(2)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}

A stirred suspension of 2, 0.683 g (1.54 mmol) in 10 mL of absoluteethanol was heated to 70° C. Methanesulfonic acid, 0.2 mL (3.08 mmol)was added dropwise to the stirred hot suspension, and heating (65-70°C.) and stirring was continued for an additional 2 minutes. Theresultant solution was left at room temperature for 4 hours. The disaltwas filtered, washed with two portions of anhydrous ethanol:ether (1:2)and then with anhydrous ether. The disalt was immediately vacuum-driedat 50-60° C. (water bath) for 2 hours to afford {[(2)(2H⁺)]²⁺·[2(CH₃SO₃⁻)]²⁻}, 0.91 g (93%), as a colorless solid; melting point 160-165° C. ¹HNMR (DMSO-d₆): δ 11.61 (s, 1H), 10.99 (brs, 1H), 8.85 (d, J=6.0 Hz, 1H),8.47 (t, J=7.2 Hz, 1H), 8.31 (s, 1H), 8.18 (d, J=6.9 Hz, 1H), ), 8.01(d, J=8.1 Hz, 1H), 7.88 (t, J=6.6 Hz, 1H), 7.83 (s, 1H), 7.44 (d, J=8.1Hz, 1H), 7.23-7.17 (m, 2H), 5.97 (s, 1H), 4.74 (m, 2H), 3.72 (m, 4H),3.59 (m, 4H), 3.46(t, J=5.7 Hz, 2H), 2.36 (s, 6H). Anal. Calcd forC₂₆H₃₃N₇O₈S₂+H₂O: C, 47.77; H, 5.40; N, 15.00; O, 22.03; S, 9.81. Found:C, 47.94; H, 5.23; N, 14.99; O, 21.87; S, 9.89.

Example 3N′-(1H-indol-3-ylmethylene)-N-methyl-N-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazine(3) dimesylate {[(3)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}

A stirred suspension of 3, 0.611 g (1.54 mmol) in 7.5 mL of absoluteethanol was heated to 70° C. Methanesulfonic acid, 0.2 mL (3.08 mmol)was added dropwise to the stirred hot suspension, and heating (65-70°C.) and stirring was continued for an additional 2 minutes. Theresultant solution was left at room temperature for 4 hours. The disaltwas filtered, washed with two portions of anhydrous ethanol:ether (1:2)and then with anhydrous ether, and then immediately vacuum-dried at50-60° C. (water bath) for 2 hours to afford {[(3)(2H⁺)]²⁺·[2(CH₃SO₃⁻)]²⁻}, 0.83 g (83%), as a light-yellow solid; melting point 130-140° C.¹H NMR (DMSO-d₆): δ 11.52 (s, 1H), 8.88 (d, J=5.1 Hz, 1H), 8.55 (t,J=8.4 Hz, 1H), 8.21 (s, 1H), 8.18 (d, J=7.2 Hz, 1H),), 8.10 (d, J=8.1Hz, 1H), 7.95 (t, J=6.6 Hz, 1H), 7.81 (d, J=2.4 Hz, 1H), 7.46 (d, J=7.2Hz, 1H), 7.22-7.16 (m, 2H), 6.41(s, 1H), 4.72 (t, J=6.0 Hz, 2H), 3.72(m, 4H), 3.51-3.48 (m, 7H), 3.47(t, J=5.1 Hz, 2H), 2.36 (s, 6H). Anal.Calcd for C₂₇H₃₅N₇O₈S₂+1.25H₂O: C, 48.31; H, 5.48; N, 14.61; O, 22.05;S, 9.55. Found: C, 48.17; H, 5.41; N, 14.34; O, 22.40; S, 9.68.

Example 4N-(3-Methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-morpholin-4-yl-ethoxy)-pyrimidin-4-yl]-hydrazine(4) dimesylate {[(4)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}

A stirred suspension of 4, 0.263 g (0.62 mmol) in 2 mL of toluene washeated to 70° C. until all solids were dissolved. Methanesulfonic acid,0.08 mL (1.24 mmol) was added dropwise to the stirred hot solution, andheating (65-70° C.) and stiffing was continued for an additional 2minutes. The resultant solution was left at room temperature for 4hours. The disalt was filtered, washed with two portions of anhydrousethanol:ether (1:2) and then with anhydrous ether, and immediatelyvacuum-dried at 50-60° C. (water bath) for 2 hours to afford{[(4)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}, 0.36 g (94%). as a colorless solid,melting point 215-219° C. ¹H NMR (DMSO-d₆): δ 11.18 (brs, 1H), 9.98(brs, 1H), 8.06 (s, 1H), 7.56-7.54 (m, 2H), 7.29 (t, J=7.8 Hz, 1H), 7.20(t, J=7.5 Hz, 1H), 6.09 (s, 1H), 4.62 (m, 2H), 3.92 (m, 8H), 3.69 (m,4H), 3.60 (m, 4H), 3.20 (m, 2H), 2.37 (s, 6H), 2.34(s, 3H). Anal. Calcdfor C₂₄H₃₇N₆O₁₀S₂+H₂O: C, 45.34; H, 6.18; N, 13.22; O, 25.17; S, 10.09.Found: C, 45.49; H, 6.00; N, 13.25; O, 24.90; S, 10.36.

Example 5N-(3-Methyl-benzylidene)-N′-[4-morpholin-4-yl-6-(pyridin-2-yl-ethoxy)-[1,3,5]-triazin-2-yl]-hydrazine(5) dimesylate {[(5)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}

A stirred suspension of 5, 0.516 g (1.23 mmol) in a mixture of 2.0 mL ofabsolute ethanol and 2.0 mL of toluene was heated to 70° C. until allsolids were dissolved. Methanesulfonic acid, 0.16 mL (2.46 mmol) wasadded dropwise to the stirred hot solution, and heating (65-70° C.) andstirring was continued for an additional two 2 minutes. The resultantsolution was left at room temperature for 4 hours. The disalt wasfiltered, washed with two portions of anhydrous ethanol:ether (1:2) andthen with anhydrous ether, and immediately vacuum-dried at 50-60° C.(water bath) for 2 hours to afford {[(5)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}, 0.707g (94%), as a colorless solid; melting point 143-147° C. ¹H NMR(DMSO-d₆): δ 11.78 (brs, 1H), 11.18 (brs, 1H), 8.87 (d, J=5.7 Hz, 1H),8.52 (t, J=8.4 Hz, 1H), 8.10-8.06 (m, 2H), 7.93 (t, J=6.3 Hz, 1H), 7.53(m, 2H), 7.32 (t, J=6.3 Hz, 1H), 6.09 (s, 1H), 4.62 (m, 2H), 3.92 (m,8H), 3.69 (m, 4H), 3.60 (m, 4H), 3.20 (m, 2H), 2.37 (s, 6H), 2.34(s,3H). Anal. Calcd for C₂₄H₃₃N₇O₈S₂+2.25H₂O+0.25 MeSO₃H: C, 43.35; H,5.74; N, 14.62; S, 10.52. Found: C, 43.32; H, 5.55; N, 14.51; S, 10.31.

Example 6N-(1H-Indol-3-ylmethylene)-N′-[4-morpholin-4-yl-6-(2-pyridin-2-yl-ethoxy)-[1,3,5]-triazin-2-yl]-hydrazine(6) dimesylate {[(6)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}

A stirred suspension of 6, 1.37 g (3.08 mmol) in 20 mL of absoluteethanol was heated to 70° C. Methanesulfonic acid, 0.4 mL (6.16 mmol)was added dropwise to the stirred hot suspension, and heating (65-70°C.) and stirring was continued for an additional 2 minutes. Theresultant solution was left at room temperature for 4 hours. The disaltwas filtered, washed with two portions of anhydrous ethanol:ether (1)and then with anhydrous ether, and immediately vacuum-dried at 50-60° C.(water bath) for 2 hours to afford {[(6)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}, 1.61g (82%), as an off-white solid; melting point 158-161° C. ¹H NMR(DMSO-d₆): δ 11.60-11.46 (brm, 2H), 8.87 (d, J=3.8 Hz, 1H), 8.50 (m,1H), 8.35-8.30 (m, 2H), 8.07 (d, J=7.2 Hz, 1H), 7.92 (m, 1H), 7.81-7.78(m, 1H), 7.42 (d, J=8.1 Hz, 1H), 7.12 (m, 1H), 7.09 (t, J=7.5 Hz, 1H),4.86 (m, 1H), 4.68 (m, 1H), 3.74 (m, 4H), 3.66 (m, 4H), 3.47-3.43(m,2H), 2.35 (s, 6H). Anal. Calcd for C₂₅H₃₁N₈O₈S₂+1.25H₂O: C, 4845.62; H,5.13; N, 17.02; O, 22.48; S, 9.74. Found: C, 45.54; H, 5.16; N, 16.77;O, 22.51; S, 10.02.

Example 7[7-Morpholin-4-yl-5-(2-pyridin-2-yl-ethoxy)-3H-[1,2,4]-triazolo[1,5-a][1,3,5]-triazin-2-yl]-m-tolyl-amine(7) dimesylate {[(7)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}

A stirred suspension of 7, 0.332 g (0.15 mmol) in 2 mL of absoluteethanol was heated to 70° C. Methanesulfonic acid, 0.02 mL (0.3 mmol)was added dropwise to the stirred hot suspension, and heating (65-70°C.) and stirring continued for an additional 2 minutes. The resultantsolution was cooled and then concentrated to leave the minimum amount ofsolvent. To the resultant solution, toluene, 1 mL, was added, and thesolvent gently distilled until precipitation started, or until about 0.5mL of ethanol remained. The mixture was left at room temperature for 4hours. The disalt was filtered, washed with two portions of anhydrousethanol:ether (1:2) and then with anhydrous ether, and immediatelyvacuum-dried at 50-60° C. (water bath) for 2 hours to afford{[(7)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}; 0.45 g (94%), as an off-white solid,melting point 137-141° C. NMR (DMSO-d₆): δ 9.58 (brs, 1H), 8.84 (d,J=5.1 Hz, 1H), 8.47 (t, J=7.1 Hz, 1H), 8.02 (d, J=7.8 Hz, 1H), 7.89 (t,J=7.6 Hz, 1H), 7.41 (s, 1H), 7.37 (d, J=8.4 Hz, 1H), 7.18 (t, J=7.8 Hz,1H), 6.79 (t, J=7.2 Hz, 1H), 4.70 (m, 2H), 3.99 (m, 4H), 3.73 (m, 4H),3.45 (t, J=5.7 Hz, 2H), 2.37 (s, 6H), 2.27 (s, 3H).

Example 8N-(3-Methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazine(1) dimethanesulfonate (dimesylate) {[(1)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}

Procedure 1: A stirred suspension of 1, 11.3 g (27 mmol) in 80 mL ofabsolute ethanol was heated to 70° C. until all solids were dissolved.Methanesulfonic acid, 3.5 mL (54 mmol) was added dropwise to the stirredhot solution, and heating (65-70° C.) and stiffing was continued for 2minutes. The resultant solution was left at room temperature for 2 hours(precipitation started soon, slowly, within ca. 5 minutes). The disaltwas filtered, washed with two portions of anhydrous ethanol:ether (1:2)(18, 36 mL) and then with 26 mL of anhydrous ether, and immediatelyvacuum-dried at 50-60° C. (water bath) for 2 hours, followed byvacuum-drying over a desiccant for 2 hours to afford{[(1)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)]²⁻}, 16.1 g (97.6%), as a colorless solid,melting point 194-195° C. ¹H NMR (DMSO-d₆): δ 8.86 (d, J=5.4 Hz, 1H),8.51 (t, J=7.8 Hz, 1H), 8.05 (m, 1H), 8.04 (s, 1H), 7.92 (t, J=6.0 Hz,1H), 7.59-7.55 (m, 2H), 7.31 (t, J=7.5 Hz, 1H), 7.21 (d, J=7.5 Hz, 1H),6.02 (s, 1H), 4.71 (m, 2H), 3.68 (m, 4H), 3.58 (m, 4H), 3.47 (t, J=6.9Hz, 2H), 2.37 (s, 3H), 2.36 (s, 3H), 2.34 (s, 3H). Anal. Calcd forC₂₅H₃₄N₆O₈S₂: C, 49.17; H, 5.61; N, 13.76; O, 20.96; S, 10.50. Found: C,49.24; H, 5.63; N, 13.63; O, 20.80; S, 10.56. % H₂O was 0.11%.

Procedure 2: Compound 1, (548.1 mg, 1.3 mmol) was dissolved in a mixtureof toluene (6 mL) and absolute ethanol (0.5 mL) by stirring at roomtemperature. To the stirred solution methanesulfonic acid, (0.17 mL, 2eq) was added. Precipitation of the salt started soon after addition ofthe methanesulfonic acid. The slurry was stirred for 2 hours to completeprecipitation, the solid was filtered out, washed two times withabsolute ethanol, and vacuum-dried to remove the residues of solvents.N-(3-methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazinedimethanesulfonate was obtained in 98%, with ¹H NMR, m.p., and elementalanalysis as given above.

This procedure is less exothermic than procedure I. Procedure II mayalternatively be carried out by dissolving the methanesulfonic acid inabsolute ethanol before adding it to the toluene/ethanol solution of 1.Methanol and 1-butanol may be substituted for ethanol in Procedure II.

Procedure III. Compound 1, (548.1 mg, 1.3 mmol) was dissolved inabsolute ethanol (8 mL) by heating to 75° C. To the stirred solution atthat temperature methanesulfonic acid, (0.17 mL, 2 eq) in water (from0.17 to 0.2 mL) was added. After addition, the stirring turned off, andthe resulted clear solution was slowly cooled to room temperature(within 3.5-4 hours). Precipitation of the salt started at around 35° C.Reaction mixture was left overnight to complete precipitation. The solidwas filtered out, washed two times with absolute ethanol, then once withethyl ether and vacuum-dried to remove the residues of solvents.N-(3-methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazinedimethanesulfonate was obtained in 85%, with ¹H NMR, m.p., and elementalanalysis as given above. Procedure III yielded agglomerated material.

Procedure IV. Compound 1, (548.1 mg, 1.3 mmol) was dissolved in DMF (1mL) by heating to 71° C. To the stirred hot solution methanesulfonicacid, (0.17 mL, 2 eq) in water (0.2 mL) was added. After the additionwas complete, the stiffing and heating were turned off, and the resultedclear solution was allowed to cool to room temperature (within 3.5-4hours). Precipitation of the salt started at around 43° C. Reactionmixture was left overnight to complete precipitation. The solid wasfiltered out, washed two times with absolute ethanol, then once withethyl ether and vacuum-dried to remove the residues of solvents.N-(3-methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazinedimethanesulfonate was obtained in 85%, with ¹H NMR, m.p., and elementalanalysis as given above.

Recrystallization Procedure I.N-(3-methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazinedimethanesulfonate (6 g, 9.8 mmol)) in 2.8 mL of purified water wasstirred at 50° C. to achieve a clear solution. To the solution absoluteethanol, 28 mL, was added, and the resulted solution was allowed to coolto room temperature. without stirring, and left overnight to completethe precipitation. The agglomerated solid was filtered out, washed twicewith ethanol (10+10 mL), then once with ethyl ether (25 mL), andvacuum-dried to remove the residues of solvents. Yield 78%.

Recrystallization Procedure II.N-(3-methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazinedimethanesulfonate (7.4 g, 12 mmol)) was dissolved in a mixture ofpurified water (5.4 mL) and 1-butanol (208.5 mL) by stiffing at 70° C.After a clear solution was achieved, heating and stiffing were turnedoff, and solution was left overnight for precipitation to be completed.The agglomerated solid was filtered out, washed twice with ethanol(20+20 mL), then once with ethyl ether (30 mL), and vacuum-dried toremove the residues of solvents. Yield 76%.

Example 9N-(3-Methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazine(1) dihydrogenchloride (dichloride) {[(1)(2H⁺)]²⁺·[2(Cl⁻)]²⁻}

Procedure I. A stirred suspension of 1, 7.7 g (18.4 mmol) in 65 mL ofabsolute ethanol was heated to 65-70° C. until everything dissolved.Into a stirred warm solution dry HCl gas was bubbled slowly. Someprecipitate started to form; then flow of gas was increased and a clearsolution was achieved. Bubbling continued until solution turned paleagain, and precipitation started; the resulted solution was left forprecipitation to be completed for 2 hours. The salt was filtered out,washed with two portions of anhydrous ethanol:ether 1:2 mixture (12:24mL), then with 16 mL of anhydrous ether and immediately vacuum-dried at50-60° C. (water bath) for 2 hours, followed by vacuum-drying over thedesiccant for 2 hours to afford 1 dihydrochloride, 8.3 (88%), colorlessto off-white solid, melting point 171-173° C. (decompos).

¹H NMR (DMSO-d₆): δ 8.85 (d, J=5.4 Hz, 1H), 8.50 (td, J=8.1 and 1.5 Hz,1H), 8.09 (s, 1H), 8.04 (d, J=8.4 Hz, 1H), 7.91 (t, J=7.2 Hz, 1H),7.59-7.55 (m, 2H), 7.31 (t, J=7.5 Hz, 1H), 7.21 (d, J=7.8 Hz, 1H), 6.01(s, 1H), 4.79 (m, 2H), 3.68 (m, 4H), 3.59 (m, 6H), 2.34 (s, 3H). Anal.Calcd for C₂₃H₂₈Cl₂N₆O₂+1H₂O: C, 54.23; H, 5.94; Cl, 13.92; N, 16.50.Found: C, 54.08; H, 5.93; Cl, 13.81; N, 16.26.

Procedure II. A stirred suspension of 1, 2.51 g (6 mmol) in 30 mL ofabsolute ethanol was heated to 65-70° C. until everything dissolved. Thestirred solution was allowed to cool down to room temperature, and a 2.0M solution of hydrogen chloride in ether, 6.01 mL (12 mmol), was addeddropwise. The stirring continued for 2 min, and the resulted solutionwas left at room temperature for precipitation for 2 hours. The salt wasfiltered out, washed with two portions of anhydrous ethanol:ether 1:2mixture (1.8:3.6 mL), then with 3 mL of anhydrous ether and immediatelyvacuum-dried at 50-60° C. (water bath) for 2 hours, followed byvacuum-drying over the desiccant for 2 hours to afford 1dihydrochloride, 2.2 g (72%), colorless to off-white solid.

¹H NMR (DMSO-d₆): δ 8.85 (d, J=5.4 Hz, 1H), 8.53 (td, J=8.1 and 1.5 Hz,1H), 8.12 (s, 1H), 8.07 (d, J=8.4 Hz, 1H), 7.93 (t, J=7.2 Hz, 1H), 7.60(m, 2H), 7.31 (t, J=7.5 Hz, 1H), 7.21 (d, J=7.8 Hz, 1H), 6.02 (s, 1H),4.79 (m, 2H), 3.68 (m, 4H), 3.59 (m, 6H), 2.34 (s, 3H).

Example 10N-(3-Methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazine(1) dinitrate {[(1)(2H⁺)]²⁺·[2(NO₃ ⁻)]²⁻}

A stirred suspension of 1, 1.13 g (2.7 mmol) in a mixture of 9.3 mL ofabsolute ethanol was heated to 70° C. until all solids were dissolved.To the stirred, hot solution was added dropwise a solution of 70% nitricacid, 0.49 g (5.4 mmol) in 3.7 mL of absolute ethanol. Stirring wascontinued for an additional 2 minutes. The resultant solution was leftat room temperature for 2 hours. The disalt was filtered, washed withtwo portions of anhydrous ethanol:ether (1:2) (1.8:3.6 mL) and then with2.6 mL of anhydrous ether, and immediately vacuum-dried at 50-60° C.(water bath) for 2 hours, followed by vacuum-drying over the desiccantfor 2 hours to afford {[(1)(2H⁺)]²⁺·[2(NO₃ ⁻)]²⁻}, 1.41 g (97%), as acolorless solid; melting point 144-145° C. ¹H NMR (DMSO-d₆): δ 8.86 (d,J=5.4 Hz, 1H), 8.50 (td, J=8.1 and 1.5 Hz, 1H), 8.08 (d, J=8.4 Hz, 1H),8.05 (s, 1H), 7.92 (t, J=7.2 Hz, 1H), 7.60-7.56 (m, 2H), 7.31 (t, J=7.5Hz, 1H), 7.19 (d, J=7.8 Hz, 1H), 6.01 (s, 1H), 4.71 (m, 2H), 3.70 (m,4H), 3.59 (m, 4H), 3.45 (m, 2H), 2.34 (s, 3H). Anal. Calcd forC₂₃H₂₈N₈O₈: C, 50.73; H, 5.18; N, 20.58; O, 23.51. Found: C, 50.78; H,5.08; N, 20.35; O, 23.79.

Compound 1 can occlude solvents rapidly; therefore disalts prepared fromcompound 1 were vacuum dried immediately after isolation.

¹H NMR spectra of compound 1 disalts shows that the signals of twopyridine protons significantly shifted to the lower field; from 8.53 and7.78 ppm for corresponding protons in the parent compound (1) to 8.85and 8.40 ppm for the protons in disalts.

Satisfactory elemental analyses were obtained for disalts. In the caseof dihydrogenhalogenides, however, water content was found to be about3%.

Table 1 summarizes the behaviour of representative compounds 1-14 underdisalt formation conditions described herein.

TABLE 1 Compound Acid PKa Result 1 CH₃SO₃H −1.2 Formed disalt,{[(1)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 1 HCl −4.5 Formed disalt,{[(1)(2H⁺)]²⁺•[2(Cl⁻)]²⁻} 1 HBr −7 Formed disalt,{[(1)(2H⁺)]²⁺•[2(Br⁻)]²⁻} 1 HNO3 −1.32 Formed disalt,{[(1)(2H⁺)]²⁺•[2(NO₃ ⁻)]²⁻} 1 H₂SO₄ −3 Formed disalt,{[(1)(2H⁺)]²⁺•[2(HSO₄ ⁻)]²⁻} 1 PhSO₃H 0.7 Formed disalt,{[(1)(2H⁺)]²⁺•[2(PhSO₃ ⁻)]²⁻} 1 p-TolSO₃H −1.34 Formed disalt,{[(1)(2H⁺)]²⁺•[2(p-TolSO₃ ⁻)]²⁻} 2 CH₃SO₃H −1.2 Formed disalt,{[(2)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 3 CH₃SO₃H −1.2 Formed disalt,{[(3)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 4 CH₃SO₃H −1.2 Formed disalt,{[(4)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 5 CH₃SO₃H −1.2 Formed disalt,{[(5)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 6 CH₃SO₃H −1.2 Formed disalt,{[(6)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 7 CH₃SO₃H −1.2 Formed disalt,{[(7)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 8 CH₃SO₃H −1.2 Isolated mixture ofdisalt, {[(8)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻}, and monosalt,{[(8)(1H⁺)]¹⁺•[1(CH₃SO₃ ⁻)]¹⁻} 9 CH₃SO₃H −1.2 Formed disalt,{[(9)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 10 CH₃SO₃H −1.2 Formed disalt,{[(10)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 11 CH₃SO₃H −1.2 Formed mono salt only,{[(11)(1H⁺)]¹⁺•[1(CH₃SO₃ ⁻)]¹⁻} 12 CH₃SO₃H −1.2 Formed mono salt only,{[(12)(1H⁺)]¹⁺•[1(CH₃SO₃ ⁻)]¹⁻} 13 CH₃SO₃H −1.2 Formed mono salt only,{[(13)(1H⁺)]²⁺•[1(CH₃SO₃ ⁻)]¹⁻} 14 CH₃SO₃H −1.2 No salt formationobserved

Disalt, {[(1)(2H⁺)]²⁺·[2(Cl⁻)]²⁻}, was determined to be soluble in waterat a concentration of 5 mg/0.1 mL. This salt further contains about 3%water.

Disalt, {[(1)(2H⁺)]²⁺·[2(Br⁻)]²⁻}, was determined to have a meltingpoint of 153-155° C. and to be soluble in water at a concentration of 5mg/0.1 mL. This salt further contains about 3.1% water.

Disalt, {[(1)(2H⁺)]²⁺·[2(NO₃ ⁻)]²⁻}, was determined to have a meltingpoint of 142-143° C. and to be insoluble in water at a concentration of5 mg/0.1 mL and partially soluble in water at a concentrations of 5mg/0.2 mL and 5 mg/0.25 mL.

Disalt, {[(¹)(²H⁺)]²⁺·[2(HSO₄ ⁻)]²⁻} was determined to have a meltingpoint of 127-130° C. and was soluble in water at concentrations of 5mg/0.1 mL.

Disalt, {[(1)(2H⁺)]²⁺·[2(PhSO₃ ⁻)]²⁻} was determined to have a meltingof 194-195° C. and to be insoluble in water at a concentration of 5mg/0.1 mL and partially soluble in water at a concentrations of 5 mg/0.2mL and 5 mg/0.25 mL. This salt further contains about 0.11% water.

Disalt, {[(1)(2H⁺)]²⁺·[2(p-TolSO₃ ⁻)]²⁻} was determined to have amelting point of 179-181° C. and to be insoluble in water at aconcentration of 5 mg/0.1 mL, partially soluble in water at aconcentration of 5 mg/0.2 mL, and soluble in water with heating at aconcentration of 5 mg/0.25 mL.

Example 11 Mono-SaltsN-(3-Methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazine1 methanesulfonate (mesylate)

A stirred suspension of 1, 2.51 g (6 mmol) in a mixture of 7.5 mL ofabsolute ethanol and 10 mL of toluene was heated to 70° C. untileverything dissolved. To the stirred hot solution a methanesulfonicacid, 0.39 mL (6 mmol) was added dropwise, heating and stiffingcontinued for 1 min, and the resulted solution was left at roomtemperature for precipitation (precipitation started soon, slowly) for 2hours. The salt was filtered out, washed with two portions of absoluteethanol (4 mL), and vacuum-dried at 50-60° C. (water bath) for 2 hoursto afford 1 mesylate 2.9 g (94%), colorless solid, melting point162-163° C.

¹H NMR (DMSO-d₆): δ 11.07 (brs, 1H), 8.81 (d, J=5.7 Hz, 1H), 8.39 (t,J=7.8 Hz, 1H), 8.02 (s, 1H), 7.93 (d, J=8.1 Hz, 1H), 7.81 (t, J=6.6 Hz,1H), 7.54-7.51 (m, 2H), 7.30 (t, J=7.8 Hz, 1H), 7.19 (d, J=7.5 Hz, 1H),6.04 (s, 1H), 4.65 (t, J=6.0 Hz, 2H), 3.67 (m, 4H), 3.55 (m, 4H),3.39(m, 2H), 2.34 (s, 6H). Anal. Calcd for C₂₄H₃₀N₆O₅S+⅔H₂O+⅔EtOH: C,54.60; H, 6.39; N, 15.08; S, 5.75. Found: C, 54.86; H, 6.14; N, 14.78;S, 5.99.

A stirred suspension of 1, 2.51 g (6 mmol) in 30 mL of absolute ethanolwas heated to 65-70° C. until everything dissolved. To the stirred hotsolution a methanesulfonic acid, 0.39 mL (6 mmol) was added dropwise,and stirring continued for 1 min, and the resulted solution was left atroom temperature for precipitation (precipitation started in 2 min) for45 min. The salt was filtered out, washed with of absolute ethanol (4mL), than with anhydrous ether (4 ml), and vacuum-dried at 50-60° C.(water bath) for 2 hours to afford 1 mesylate 3.08 g (99%), colorlesssolid, melting point 162-163° C.

N-(3-Methyl-benzylidene)-N′-[6-morpholin-4-yl-2-(2-pyridin-2-yl-ethoxy)-pyrimidin-4-yl]-hydrazine(1) hydrochloride

A stirred suspension of 1, 2.51 g (6 mmol) in 30 mL of absolute ethanolwas heated to 65-70° C. until everything dissolved. The stirred solutionwas allowed to cool down to room temperature, and a 2.0 M solution ofhydrogen chloride in ether, 3.01 mL (6 mmol), was added dropwise. Thestiffing continued for 2 min, and the resulted solution was left at roomtemperature for precipitation for 2 hours. The salt was filtered out,washed with two portions of anhydrous ethanol:ether 1:2 mixture (1.8:3.6mL), then with 3 mL of anhydrous ether and immediately vacuum-dried at50-60° C. (water bath) for 1 hour to afford 1 hydrochloride, 2.04 g(75%), colorless to off-white solid, melting point 176-178° C.

¹H NMR (DMSO-d₆): δ 11.04 (brs, 1H), 8.77 (d, J=5.1 Hz, 1H), 8.34 (t,J=8.1 and 1.5 Hz, 1H), 8.02 (s, 1H), 7.88 (d, J=8.1 Hz, 1H), 7.77 (t,J=7.2 Hz, 1H), 7.54-7.51 (m, 2H), 7.30 (t, J=7.6 Hz, 1H), 7.18 (d, J=7.2Hz, 1H), 6.04 (s, 1H), 4.64 (t, J=5.7 Hz, 2H), 3.67 (m, 4H), 3.54 (m,4H), 3.42 (m, 2H), 2.34 (s, 3H). Anal. Calcd for C₂₃H₂₇ClN₆O₂+¼H₂O: C,60.12; H, 6.03; N, 18.29; Cl, 7.72. Found: C, 60.39; H, 6.05; N, 18.03;Cl, 7.87.

Example 12 Solubility Determination

The compound was weighed, and water was added to it such that the ratioof the sample and water of the mixture would be ˜100 mg/mL. The mixturewas then shaken using a Vortex-Genie2 shaker (vortexed) and sonicated(˜5-10 min at ˜50° C.). If a clear solution has been obtained, morecompound was added to the solution, and a mixture was vortexed andsonicated until a homogeneous suspension was achieved. The suspensionwas then centrifuged at10,000 rpm for about 10 min. The supernatant wastaken out and diluted with 50/50 (v/v) EtOH/water, then the sample wasanalyzed by HPLC to determine the concentration.

Instrumentation:

The HPLC system consisted of the HP 1100 Model (Agilent, Wilmington,Del.) equipped with a model 1100 quaternary pump, a model 1100autosampler, a model 1100 Diode-Array Detector for UV detection at 320nm. The HPLC analysis was performed using an isocratic mobile phaseconsisting of acetonitrile-water containing 10 mM NH₄OAc. Mobile phaseswere degassed and filtered through a solvent filtration apparatus andpumped at a constant rate of 1.0 mL/min. The separation was made on anXTerra MS C18 analytical column, 4.6 mm i.d.×150 mm, (Waters Corp.,Milford, Mass., USA) fitted with a precolumn filter (XTerra MS C18, 3.9mm×20 mm). The column was maintained at 30° C. Data acquisition andinstrument setting were controlled using HP Chemstation software (ver.8.03).

Table 2 summarizes the solubilities of various salts.

TABLE 2 Salt Solubility (mg/mL) {[(1)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 831 +/−5  {[(1)(1H⁺)]¹⁺•[1(CH₃SO₃ ⁻)]¹⁻} 52.0 +/− 0.1 {[(1)(2H⁺)]²⁺•[2(Cl⁻)]²⁻}213 +/− 5  {[(1)(2H⁺)]¹⁺•[7(Cl⁻)]¹⁻} 12.6 +/− 0.4{[(1)(2H⁺)]²⁺•[2(Br⁻)]²⁻} 246 +/− 3  {[(1)(2H⁺)]²⁺•[2(NO₃ ⁻)]²⁻} 83.4+/− 0.2 {[(1)(2H⁺)]²⁺•[2(p-TolSO₃ ⁻)]²⁻} 12.1 +/− 0.1{[(1)(2H⁺)]²⁺•[2(PhSO₃ ⁻)]²⁻} 14.6 +/− 0.0 {[(3)(2H⁺)]²⁺•[2(CH₃SO₃⁻)]²⁻} 306 +/− 14 {[(2)(2H⁺)]²⁺•[2(CH₃SO₃ ⁻)]²⁻} 454 +/− 5 

The solubility of disalt {[(1)(2H⁺)]²⁺·[2(CH₃SO₃ ⁻)²⁻} was,unexpectedly, more than ten times greater than the solubility of thecorresponding monosalt, {[(1)(1H⁺)]¹⁺·[1(CH₃SO₃ ⁻)]¹⁻]. The solubilitiesof disalts of compound 1 having inorganic anionic charge balancingmoieties were higher than the solubility of monosalt,{[(1)(1H⁺)]¹⁺·[1(CH₃SO₃ ⁻)]¹⁻}. The solubilities of disalts of compounds2 or 3 having mesylate anionic charge balancing moities were also higherthan the solubility of monosalt, {[(1)(1H⁺)]¹⁺·[1(CH₃SO₃ ⁻)]¹⁻}.Moreover, the disalts are less colored in appearance (indicating greaterstability, less prone to decomposition) and appear to have lessersensitivity to exposure to light (i.e., better light stability).

A number of embodiments of the invention have been described.Nevertheless, it will be understood that various modifications may bemade without departing from the spirit and scope of the invention.Accordingly, other embodiments are within the scope of the followingclaim

1.-55. (canceled)
 56. A method for treating an interleukin-12overproduction-related disorder, comprising administering to a subjectin need thereof an effective amount of a disalt represented by formula:

each M⁻ is a conjugate base of a Bronsted acid. 57-62. (canceled) 63.The method of claim 56, wherein the disorder is selected from the groupconsisting of multiple sclerosis, sepsis, myasthenia gravis, autoimmuneneuropathies, Guillain-Barré syndrome, autoimmune uveitis, autoimmunehemolytic anemia, pernicious anemia, autoimmune thrombocopenia, temporalarteritis, anti-phospholipid syndrome, vasculitides, Wegener'sgranulomatosis, Behcet's disease, psoriasis, psoriatic arthritis,dermatitis herpetiformis, pemphigus vulgaris, vitiligo, Crohn's disease,ulcerative colitis, interstitial pulmonary fibrosis, myelofibrosis,hepatic fibrosis, myocarditis, thyroditis, primary biliary cirrhosis,autoimmune hepatitis, immune-mediated diabetes mellitus, Grave'sdisease, Hashimoto's thyroiditis, autoimmune oophoritis and orchitis,autoimmune disease of the adrenal gland; rheumatoid arthritis, juvenilerheumatoid arthritis, systemic lupus erythematosus, scleroderma,polymyositis, dermatomyositis, spondyloarthropathies, ankylosingspondylitis, Sjogren's syndrome and graft-versus-host disease.
 64. Themethod of claim 63, wherein the disorder is rheumatoid arthritis,sepsis, Crohn's disease, multiple sclerosis, psoriasis, orimmune-mediated diabetes mellitus.
 65. A method of treating orpreventing an inflammatory disorder in a subject in need of suchtreatment, comprising administering to the subject an effective amountof a disalt represented by formula:

each M⁻ is a conjugate base of a Bronsted acid.
 66. A method of treatingan immune disease in a subject in need of such treatment, comprisingadministering to the subject an effective amount of a disalt representedby formula:

each M⁻ is a conjugate base of a Bronsted acid.
 67. The method accordingto claim 56, wherein M⁻ in the disalt is the conjugate base of aBronsted acid selected from the group consisting of hydrochloric acid,nitric acid, sulfuric acid, hydrobromic acid, hydroiodic acid,perchloric acid, phosphoric acid, alkylsulfonic acids, arylsulfonicacids, halogenated alkylsulfonic acids, halogenated acetic acids, picricacid, oxalic acid, citric acid, formic acid, ascorbic acid and benzoicacid.
 68. The method of claim 67, wherein M⁻ is methanesulfonate,bromide, or chloride.
 69. The method according to claim 65, wherein M⁻in the disalt is the conjugate base of a Bronsted acid selected from thegroup consisting of hydrochloric acid, nitric acid, sulfuric acid,hydrobromic acid, hydroiodic acid, perchloric acid, phosphoric acid,alkylsulfonic acids, arylsulfonic acids, halogenated alkylsulfonicacids, halogenated acetic acids, picric acid, oxalic acid, citric acid,formic acid, ascorbic acid and benzoic acid.
 70. The method of claim 69,wherein M⁻ is methanesulfonate, bromide, or chloride.
 71. The methodaccording to claim 66, wherein M⁻ in the disalt is the conjugate base ofa Bronsted acid selected from the group consisting of hydrochloric acid,nitric acid, sulfuric acid, hydrobromic acid, hydroiodic acid,perchloric acid, phosphoric acid, alkylsulfonic acids, arylsulfonicacids, halogenated alkylsulfonic acids, halogenated acetic acids, picricacid, oxalic acid, citric acid, formic acid, ascorbic acid and benzoicacid.
 72. The method of claim 71, wherein M⁻ is methanesulfonate,bromide, or chloride.